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  • 99
    Thermo Fisher hygromycin
    Molecular and biochemical analysis of transgenic rice lines co-overexpressing OsGS1;1 and OsGS2 . (A) PCR amplification of <t>hygromycin</t> phosphotransferase ( hpt ), OsGS1;1 and OSGS2 genes using specific primers in wild type ( wt ), null segregant ( ns ), and five positive T 2 transgenic lines (L1-L5). M: 1Kb DNA ladder (+): positive PCR control (pMDC99) and (–) water blank. (B) Southern blot analysis of wt and five T 2 transgenic lines (L1, L2, L3, L4, and L5), probed with hpt gene probe showing single copy insertion. (C) Semi quantitative RT-PCR showing overexpression of OsGS1;1 and OsGS2 in transgenic lines (L1, L4, and L5) as compared to wt . The rice eEF1α gene was used as a reference gene and rRNA was used as loading control. (D ; top panel) Immunoblot analysis of three transgenic rice lines (L1, L4, and L5) and wt using a recombinant antibody which detects both OsGS1;1 and OsGS2 isoforms (black lines separate spliced regions from same blot) ( D ; bottom panel) Coomassie blue stained Rubisco large subunit (RubL) was used as loading control. (E) Total GS activity of three transgenic rice lines (L1, L4, and L5) in comparison to wt ). One unit of GS activity represents 1.0 μmol of γ-glutamylhydroxamate produced in 20 min. Asterisks above bars indicate significant differences from wt (* at p ≤ 0.05 and ** at p ≤ 0.01).
    Hygromycin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6630 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Amresco hygromycin
    Mycobacterium tuberculosis cultured on 7H10 solid medium in the absence of the inducer ( Section 1 and Section 3 ) and the presence of 0.1 µg/mL pristinamycin ( Section 2 and Section 4 ). Plate B contained 100 µg/mL <t>hygromycin.</t> ( A ) MTB H37Rv bacteria formed a lawn of colonies on the medium and were dependent on the inducer for survival; ( B ) MTB:: asadh bacteria formed colonies only on the medium containing pristinamycin. The experiments were repeated three times, and typical images are shown.
    Hygromycin, supplied by Amresco, used in various techniques. Bioz Stars score: 93/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Avantor hygromycin
    Genotypic and phenotypic analysis of genetically manipulated parasites. (A, B, and C) Genomic DNA from the following parasite strains was used as the template: lanes 1, wild type; lanes 2, ARG/arg heterozygote; lanes 3, Δ arg mutant 1; lanes 4, Δ arg mutant 2; lanes 5, Δ arg [ ARG ] mutant; lanes 6, Δ arg [ arg ΔAKL] mutant. (A) Primers designed to encompass the coding region were used to amplify the ARG gene. (B) The forward primer sequence was located in a region upstream of the 5′ flanking sequence (upstream of the targeting construct), and the reverse primer sequence was located within the <t>hygromycin</t> resistance gene. (C) The same sense primer as that used for panel B was used, and the antisense primer sequence was located within the phleomycin resistance gene. (D) Western blot analyses were performed with cell extracts prepared from wild-type, ARG/arg heterozygote, Δ arg mutant 1, Δ arg mutant 2, Δ arg [ ARG ], and Δ arg [ arg ΔAKL] parasites. Parasites were fractioned by SDS-PAGE and the blot probed with polyclonal antibodies against L. mexicana ARG and a commercial monoclonal antibody that recognizes tubulin. The tubulin antibody was employed to verify equal loading of protein on all lanes.
    Hygromycin, supplied by Avantor, used in various techniques. Bioz Stars score: 93/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    BioShop hygromycin
    Genotypic and phenotypic analysis of genetically manipulated parasites. (A, B, and C) Genomic DNA from the following parasite strains was used as the template: lanes 1, wild type; lanes 2, ARG/arg heterozygote; lanes 3, Δ arg mutant 1; lanes 4, Δ arg mutant 2; lanes 5, Δ arg [ ARG ] mutant; lanes 6, Δ arg [ arg ΔAKL] mutant. (A) Primers designed to encompass the coding region were used to amplify the ARG gene. (B) The forward primer sequence was located in a region upstream of the 5′ flanking sequence (upstream of the targeting construct), and the reverse primer sequence was located within the <t>hygromycin</t> resistance gene. (C) The same sense primer as that used for panel B was used, and the antisense primer sequence was located within the phleomycin resistance gene. (D) Western blot analyses were performed with cell extracts prepared from wild-type, ARG/arg heterozygote, Δ arg mutant 1, Δ arg mutant 2, Δ arg [ ARG ], and Δ arg [ arg ΔAKL] parasites. Parasites were fractioned by SDS-PAGE and the blot probed with polyclonal antibodies against L. mexicana ARG and a commercial monoclonal antibody that recognizes tubulin. The tubulin antibody was employed to verify equal loading of protein on all lanes.
    Hygromycin, supplied by BioShop, used in various techniques. Bioz Stars score: 93/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Boehringer Mannheim hygromycin
    Southern blot analysis of integrated HIV-1 DNA demonstrates intact viral genomes. The integrated proviral HIV-1 DNA is depicted with the LTRs flanking the gag , pol , and <t>hygromycin</t> ( hygro ) genes. The numbers 1 and 9718 indicate the terminal nucleotide residues of the HXB2 genome. Numbers below the restriction enzymes ( Kas I, Xho I, Eco RV, Cla I, Bam HI, and Hin dIII) indicate the nucleotide positions of the respective cleavage sites in the HXB2 genome. Probes are indicated by a boxed region and correspond to a 534-bp sequence from the middle of the hygro gene (A) and HXB2 sequence from positions 641 to 828 (left end) and 8604 to 8895 (right end) (B). Horizontal arrows and their associated numbers indicated the expected sizes of restriction fragments for an intact proviral genome. The Southern blot analysis proceeded in two steps, first using a hygromycin probe to detect an intact genome between the two LTRs (A). The second step used individual probes for each end (B) to check for intact sequence to within 112 bp of the left end and to within 103 bp of the right end.
    Hygromycin, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 87/100, based on 233 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Carl Roth GmbH hygromycin
    Enhancement of vitamin B 6 content in rice by heterologous expression of Arabidopsis PDX 1/ PDX 2. (a) Schematic representation of the T‐ DNA regions of the transformation vector used to generate the 35S lines. LB , Left Border; RB , Right Border; 35S , cauliflower mosaic virus 35S ( Ca MV 35S ) promoter; At PDX 1.1 , At2g38230 coding sequence; At PDX2 , At5g60540 coding sequence; hpt II , <t>hygromycin</t> phosphotransferase gene; T1, Ca MV 35S terminator; T2, Octopine synthase terminator. (b) Vitamin B 6 content in leaf sample extracts of 35S lines in the T 2 generation compared to wild‐type ( TP 309) and empty vector control (p1300). Average ± SD of four biological replicates. Student's t‐ test (p1300 versus transgenic lines and TP 309), * P
    Hygromycin, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 97/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Cellgro hygromycin
    Enhancement of vitamin B 6 content in rice by heterologous expression of Arabidopsis PDX 1/ PDX 2. (a) Schematic representation of the T‐ DNA regions of the transformation vector used to generate the 35S lines. LB , Left Border; RB , Right Border; 35S , cauliflower mosaic virus 35S ( Ca MV 35S ) promoter; At PDX 1.1 , At2g38230 coding sequence; At PDX2 , At5g60540 coding sequence; hpt II , <t>hygromycin</t> phosphotransferase gene; T1, Ca MV 35S terminator; T2, Octopine synthase terminator. (b) Vitamin B 6 content in leaf sample extracts of 35S lines in the T 2 generation compared to wild‐type ( TP 309) and empty vector control (p1300). Average ± SD of four biological replicates. Student's t‐ test (p1300 versus transgenic lines and TP 309), * P
    Hygromycin, supplied by Cellgro, used in various techniques. Bioz Stars score: 90/100, based on 115 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Duchefa hygromycin
    EM sensitivity to the selective agents. Kan: kanamycin, Hyg: <t>hygromycin.</t> Data were collected after 3 subcultures (6 weeks) and the values for the 5 embryogenic lines were averaged.
    Hygromycin, supplied by Duchefa, used in various techniques. Bioz Stars score: 94/100, based on 243 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Enzo Biochem hygromycin
    EM sensitivity to the selective agents. Kan: kanamycin, Hyg: <t>hygromycin.</t> Data were collected after 3 subcultures (6 weeks) and the values for the 5 embryogenic lines were averaged.
    Hygromycin, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 98/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    EuroClone hygromycin
    EM sensitivity to the selective agents. Kan: kanamycin, Hyg: <t>hygromycin.</t> Data were collected after 3 subcultures (6 weeks) and the values for the 5 embryogenic lines were averaged.
    Hygromycin, supplied by EuroClone, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    FUJIFILM hygromycin
    Structure and expression of the TAR1 gene. A, The TAR1 gene contains 14 exons (thick boxes) separated by 13 introns. The insertion site of the DNA cassette encoding the <t>hygromycin</t> resistance gene ( aphVII ) in tar1-1 is indicated by a black triangle. The
    Hygromycin, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 97/100, based on 373 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    InvivoGen hygromycin
    Modified Flp-In system. ( a ) A targeting vector is introduced to knock out a single allele of GSK-3β and create a host cell line with a FRT-puro cassette. Co-transfection of Flp recombinase and a gene of interest previously cloned into a vector containing a V5 epitope and FRT-hygro cassette results in homologous recombination at the FRT sites. Positive colonies are resistant to <t>hygromycin</t> and sensitive to puromycin. ( b ) Western blot analysis of transgenic myr-PDK-1 cell lines. ( c ) Membrane localization of myr-PDK-1 by immunofluorescent staining of V5; confocal, × 40 magnification.
    Hygromycin, supplied by InvivoGen, used in various techniques. Bioz Stars score: 99/100, based on 886 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Merck KGaA hygromycin
    Modified Flp-In system. ( a ) A targeting vector is introduced to knock out a single allele of GSK-3β and create a host cell line with a FRT-puro cassette. Co-transfection of Flp recombinase and a gene of interest previously cloned into a vector containing a V5 epitope and FRT-hygro cassette results in homologous recombination at the FRT sites. Positive colonies are resistant to <t>hygromycin</t> and sensitive to puromycin. ( b ) Western blot analysis of transgenic myr-PDK-1 cell lines. ( c ) Membrane localization of myr-PDK-1 by immunofluorescent staining of V5; confocal, × 40 magnification.
    Hygromycin, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 94/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Mirus Bio hygromycin
    Modified Flp-In system. ( a ) A targeting vector is introduced to knock out a single allele of GSK-3β and create a host cell line with a FRT-puro cassette. Co-transfection of Flp recombinase and a gene of interest previously cloned into a vector containing a V5 epitope and FRT-hygro cassette results in homologous recombination at the FRT sites. Positive colonies are resistant to <t>hygromycin</t> and sensitive to puromycin. ( b ) Western blot analysis of transgenic myr-PDK-1 cell lines. ( c ) Membrane localization of myr-PDK-1 by immunofluorescent staining of V5; confocal, × 40 magnification.
    Hygromycin, supplied by Mirus Bio, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PhytoTechnology Laboratories hygromycin
    Modified Flp-In system. ( a ) A targeting vector is introduced to knock out a single allele of GSK-3β and create a host cell line with a FRT-puro cassette. Co-transfection of Flp recombinase and a gene of interest previously cloned into a vector containing a V5 epitope and FRT-hygro cassette results in homologous recombination at the FRT sites. Positive colonies are resistant to <t>hygromycin</t> and sensitive to puromycin. ( b ) Western blot analysis of transgenic myr-PDK-1 cell lines. ( c ) Membrane localization of myr-PDK-1 by immunofluorescent staining of V5; confocal, × 40 magnification.
    Hygromycin, supplied by PhytoTechnology Laboratories, used in various techniques. Bioz Stars score: 93/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Roche hygromycin
    Generation of the Δ msmHr M. smegmatis strain . (A) Genomic organization of the msmHr gene locus. (Upper panel) Genes are shown as large arrows in their native orientation. Small arrows represent the forward and reverse primers used for PCR, and sizes of the amplified products are indicated. Location and orientation of the <t>hygromycin</t> cassette are also indicated (Bottom panel). No PCR product was obtained using primers 2415InL and 2415InR to amplify the coding sequences of msmHr . PCR products for the upstream and downstream regions of msmHr were amplified using the primer pairs 2415LLL/IL(R) and 2415RRR/IR(F), respectively. (B) Effect of msmHr deletion on msmeg_2414 and msmeg_2416 expression. Quantitative real-time PCR (qRT-PCR) analysis of msmHr cluster transcription. The primer pairs 2414qF/2414qR and 2416qF/2416qR were used for qRT-PCR. Results are shown as the means ± standard deviations of three replicates ( * P
    Hygromycin, supplied by Roche, used in various techniques. Bioz Stars score: 96/100, based on 1686 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology hygromycin
    Generation of the Δ msmHr M. smegmatis strain . (A) Genomic organization of the msmHr gene locus. (Upper panel) Genes are shown as large arrows in their native orientation. Small arrows represent the forward and reverse primers used for PCR, and sizes of the amplified products are indicated. Location and orientation of the <t>hygromycin</t> cassette are also indicated (Bottom panel). No PCR product was obtained using primers 2415InL and 2415InR to amplify the coding sequences of msmHr . PCR products for the upstream and downstream regions of msmHr were amplified using the primer pairs 2415LLL/IL(R) and 2415RRR/IR(F), respectively. (B) Effect of msmHr deletion on msmeg_2414 and msmeg_2416 expression. Quantitative real-time PCR (qRT-PCR) analysis of msmHr cluster transcription. The primer pairs 2414qF/2414qR and 2416qF/2416qR were used for qRT-PCR. Results are shown as the means ± standard deviations of three replicates ( * P
    Hygromycin, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Wisent Corporation hygromycin
    Generation of the Δ msmHr M. smegmatis strain . (A) Genomic organization of the msmHr gene locus. (Upper panel) Genes are shown as large arrows in their native orientation. Small arrows represent the forward and reverse primers used for PCR, and sizes of the amplified products are indicated. Location and orientation of the <t>hygromycin</t> cassette are also indicated (Bottom panel). No PCR product was obtained using primers 2415InL and 2415InR to amplify the coding sequences of msmHr . PCR products for the upstream and downstream regions of msmHr were amplified using the primer pairs 2415LLL/IL(R) and 2415RRR/IR(F), respectively. (B) Effect of msmHr deletion on msmeg_2414 and msmeg_2416 expression. Quantitative real-time PCR (qRT-PCR) analysis of msmHr cluster transcription. The primer pairs 2414qF/2414qR and 2416qF/2416qR were used for qRT-PCR. Results are shown as the means ± standard deviations of three replicates ( * P
    Hygromycin, supplied by Wisent Corporation, used in various techniques. Bioz Stars score: 93/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    AG Scientific hygromycin
    Generation of the Δ msmHr M. smegmatis strain . (A) Genomic organization of the msmHr gene locus. (Upper panel) Genes are shown as large arrows in their native orientation. Small arrows represent the forward and reverse primers used for PCR, and sizes of the amplified products are indicated. Location and orientation of the <t>hygromycin</t> cassette are also indicated (Bottom panel). No PCR product was obtained using primers 2415InL and 2415InR to amplify the coding sequences of msmHr . PCR products for the upstream and downstream regions of msmHr were amplified using the primer pairs 2415LLL/IL(R) and 2415RRR/IR(F), respectively. (B) Effect of msmHr deletion on msmeg_2414 and msmeg_2416 expression. Quantitative real-time PCR (qRT-PCR) analysis of msmHr cluster transcription. The primer pairs 2414qF/2414qR and 2416qF/2416qR were used for qRT-PCR. Results are shown as the means ± standard deviations of three replicates ( * P
    Hygromycin, supplied by AG Scientific, used in various techniques. Bioz Stars score: 95/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Becton Dickinson hygromycin
    Generation of a M. bovis BCG kasB null mutant. Maps of the kasB region in the M. bovis BCG genome and its corresponding region in the conditional mutant Δ kasB are shown on the left, with the corresponding Southern blot on the right. The regions used as porbes are indicated by solid lines with square ends while the expected bands are indicated by broken lines (with sizes indicated). hyg , <t>hygromycin</t> resistance gene from Streptomyces hygroscopicus ; res , γδ resolvase recognition sites.
    Hygromycin, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 95/100, based on 144 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Euromedex hygromycin
    Effects of the PDR1 YJM326 allele in different genetic backgrounds (A) Fitness variation of 20 isolates (left panel) in comparison with the same set of strains hybridized with YJM326 in the presence of drug. Fitness values (y-axis) correspond to the ratio between the growth in the presence of cycloheximide (YPD CHX 1µg/ml) and control media YPD. Dashed line indicates the fitness of the resistant strain YJM326. (B) Fitness variation of 20 isolates carrying empty control plasmid (pCTRL, left panel) or plasmid containing the PDR1 YJM326 allele under its native promoter (pPDR1 YJM326 , right panel). Fitness values were measured in the presence of cycloheximide (YPD CHX 1µg/ml) with <t>hygromycin</t> to maintain plasmid stability. Dashed line indicates the fitness value of YJM326 carrying the plasmid pPDR1 YJM326 for detailed comparison for effect of hybrid and plasmid in individual genetic backgrounds.
    Hygromycin, supplied by Euromedex, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Fisher Scientific hygromycin
    Effects of the PDR1 YJM326 allele in different genetic backgrounds (A) Fitness variation of 20 isolates (left panel) in comparison with the same set of strains hybridized with YJM326 in the presence of drug. Fitness values (y-axis) correspond to the ratio between the growth in the presence of cycloheximide (YPD CHX 1µg/ml) and control media YPD. Dashed line indicates the fitness of the resistant strain YJM326. (B) Fitness variation of 20 isolates carrying empty control plasmid (pCTRL, left panel) or plasmid containing the PDR1 YJM326 allele under its native promoter (pPDR1 YJM326 , right panel). Fitness values were measured in the presence of cycloheximide (YPD CHX 1µg/ml) with <t>hygromycin</t> to maintain plasmid stability. Dashed line indicates the fitness value of YJM326 carrying the plasmid pPDR1 YJM326 for detailed comparison for effect of hybrid and plasmid in individual genetic backgrounds.
    Hygromycin, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 96/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Gemini Bio hygromycin
    Effects of the PDR1 YJM326 allele in different genetic backgrounds (A) Fitness variation of 20 isolates (left panel) in comparison with the same set of strains hybridized with YJM326 in the presence of drug. Fitness values (y-axis) correspond to the ratio between the growth in the presence of cycloheximide (YPD CHX 1µg/ml) and control media YPD. Dashed line indicates the fitness of the resistant strain YJM326. (B) Fitness variation of 20 isolates carrying empty control plasmid (pCTRL, left panel) or plasmid containing the PDR1 YJM326 allele under its native promoter (pPDR1 YJM326 , right panel). Fitness values were measured in the presence of cycloheximide (YPD CHX 1µg/ml) with <t>hygromycin</t> to maintain plasmid stability. Dashed line indicates the fitness value of YJM326 carrying the plasmid pPDR1 YJM326 for detailed comparison for effect of hybrid and plasmid in individual genetic backgrounds.
    Hygromycin, supplied by Gemini Bio, used in various techniques. Bioz Stars score: 94/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    HiMedia Laboratories hygromycin
    Effects of the PDR1 YJM326 allele in different genetic backgrounds (A) Fitness variation of 20 isolates (left panel) in comparison with the same set of strains hybridized with YJM326 in the presence of drug. Fitness values (y-axis) correspond to the ratio between the growth in the presence of cycloheximide (YPD CHX 1µg/ml) and control media YPD. Dashed line indicates the fitness of the resistant strain YJM326. (B) Fitness variation of 20 isolates carrying empty control plasmid (pCTRL, left panel) or plasmid containing the PDR1 YJM326 allele under its native promoter (pPDR1 YJM326 , right panel). Fitness values were measured in the presence of cycloheximide (YPD CHX 1µg/ml) with <t>hygromycin</t> to maintain plasmid stability. Dashed line indicates the fitness value of YJM326 carrying the plasmid pPDR1 YJM326 for detailed comparison for effect of hybrid and plasmid in individual genetic backgrounds.
    Hygromycin, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Nacalai hygromycin
    Effects of the PDR1 YJM326 allele in different genetic backgrounds (A) Fitness variation of 20 isolates (left panel) in comparison with the same set of strains hybridized with YJM326 in the presence of drug. Fitness values (y-axis) correspond to the ratio between the growth in the presence of cycloheximide (YPD CHX 1µg/ml) and control media YPD. Dashed line indicates the fitness of the resistant strain YJM326. (B) Fitness variation of 20 isolates carrying empty control plasmid (pCTRL, left panel) or plasmid containing the PDR1 YJM326 allele under its native promoter (pPDR1 YJM326 , right panel). Fitness values were measured in the presence of cycloheximide (YPD CHX 1µg/ml) with <t>hygromycin</t> to maintain plasmid stability. Dashed line indicates the fitness value of YJM326 carrying the plasmid pPDR1 YJM326 for detailed comparison for effect of hybrid and plasmid in individual genetic backgrounds.
    Hygromycin, supplied by Nacalai, used in various techniques. Bioz Stars score: 97/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Formedium hygromycin
    Generation of A . benhamiae Δ hapX mutants and reconstituted strains. (A) For deletion of the hapX locus (white arrow) in the wild-type strain A . benhamiae LAU2354-2 (bottom) a DNA cassette, containing the <t>hygromycin</t> resistance gene hph (black arrow) under control of the gpd promoter (P gpd , bent arrow) together with the termination sequence fragment T trpC (filled circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used (top). (B) For reinsertion of the hapX gene into its original locus in the Δ hapX mutants a DNA cassette, containing the coding region of hapX and the neomycin resistance gene neo (lined arrow) under control of the A . benhamiae actin promoter (P ACT1 , bent arrow) together with the Candida albicans actin termination sequence fragment T ACT1 (blank circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used. (C) Southern blot of Nde I-digested genomic DNA of the wild-type strain A . benhamiae LAU2354-2, Δ hapX mutants and hapX C reconstituted strains with hapX -specific probe 1. The probes which were used for Southern analysis of the transformants are indicated by the black bars. Only the following relevant restriction sites are given in panels a and b: A, Apa I; B, Bam HI; H, Hin dIII; Nd, Nde I; Not I. The sizes of the hybridizing fragments are given on the left and their identities on the right.
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    LC Laboratories hygromycin
    Generation of A . benhamiae Δ hapX mutants and reconstituted strains. (A) For deletion of the hapX locus (white arrow) in the wild-type strain A . benhamiae LAU2354-2 (bottom) a DNA cassette, containing the <t>hygromycin</t> resistance gene hph (black arrow) under control of the gpd promoter (P gpd , bent arrow) together with the termination sequence fragment T trpC (filled circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used (top). (B) For reinsertion of the hapX gene into its original locus in the Δ hapX mutants a DNA cassette, containing the coding region of hapX and the neomycin resistance gene neo (lined arrow) under control of the A . benhamiae actin promoter (P ACT1 , bent arrow) together with the Candida albicans actin termination sequence fragment T ACT1 (blank circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used. (C) Southern blot of Nde I-digested genomic DNA of the wild-type strain A . benhamiae LAU2354-2, Δ hapX mutants and hapX C reconstituted strains with hapX -specific probe 1. The probes which were used for Southern analysis of the transformants are indicated by the black bars. Only the following relevant restriction sites are given in panels a and b: A, Apa I; B, Bam HI; H, Hin dIII; Nd, Nde I; Not I. The sizes of the hybridizing fragments are given on the left and their identities on the right.
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    Mediatech hygromycin
    Manipulation of Abl expression and activity in normal and malignant MECs. A ) Immunoblotting for c-Abl expression in NMuMG and 4T1 cells engineered to express either a scrambled (scram) or an Abl-targeting shRNA (shAbl), or empty vector <t>(pMSCV-hygromycin),</t>
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    Merck & Co hygromycin
    Manipulation of Abl expression and activity in normal and malignant MECs. A ) Immunoblotting for c-Abl expression in NMuMG and 4T1 cells engineered to express either a scrambled (scram) or an Abl-targeting shRNA (shAbl), or empty vector <t>(pMSCV-hygromycin),</t>
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    PAA Laboratories hygromycin
    Manipulation of Abl expression and activity in normal and malignant MECs. A ) Immunoblotting for c-Abl expression in NMuMG and 4T1 cells engineered to express either a scrambled (scram) or an Abl-targeting shRNA (shAbl), or empty vector <t>(pMSCV-hygromycin),</t>
    Hygromycin, supplied by PAA Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega hygromycin
    Establishment of core3 synthase-expressing HT-29 cells. A , HT-29 cells were transfected with mock and core3 synthase expression plasmids, and several <t>hygromycin-resistant</t> clones of each were isolated. A , RT-PCR results of core3 synthase expression of
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    Image Search Results


    Molecular and biochemical analysis of transgenic rice lines co-overexpressing OsGS1;1 and OsGS2 . (A) PCR amplification of hygromycin phosphotransferase ( hpt ), OsGS1;1 and OSGS2 genes using specific primers in wild type ( wt ), null segregant ( ns ), and five positive T 2 transgenic lines (L1-L5). M: 1Kb DNA ladder (+): positive PCR control (pMDC99) and (–) water blank. (B) Southern blot analysis of wt and five T 2 transgenic lines (L1, L2, L3, L4, and L5), probed with hpt gene probe showing single copy insertion. (C) Semi quantitative RT-PCR showing overexpression of OsGS1;1 and OsGS2 in transgenic lines (L1, L4, and L5) as compared to wt . The rice eEF1α gene was used as a reference gene and rRNA was used as loading control. (D ; top panel) Immunoblot analysis of three transgenic rice lines (L1, L4, and L5) and wt using a recombinant antibody which detects both OsGS1;1 and OsGS2 isoforms (black lines separate spliced regions from same blot) ( D ; bottom panel) Coomassie blue stained Rubisco large subunit (RubL) was used as loading control. (E) Total GS activity of three transgenic rice lines (L1, L4, and L5) in comparison to wt ). One unit of GS activity represents 1.0 μmol of γ-glutamylhydroxamate produced in 20 min. Asterisks above bars indicate significant differences from wt (* at p ≤ 0.05 and ** at p ≤ 0.01).

    Journal: Frontiers in Plant Science

    Article Title: Concurrent Overexpression of OsGS1;1 and OsGS2 Genes in Transgenic Rice (Oryza sativa L.): Impact on Tolerance to Abiotic Stresses

    doi: 10.3389/fpls.2018.00786

    Figure Lengend Snippet: Molecular and biochemical analysis of transgenic rice lines co-overexpressing OsGS1;1 and OsGS2 . (A) PCR amplification of hygromycin phosphotransferase ( hpt ), OsGS1;1 and OSGS2 genes using specific primers in wild type ( wt ), null segregant ( ns ), and five positive T 2 transgenic lines (L1-L5). M: 1Kb DNA ladder (+): positive PCR control (pMDC99) and (–) water blank. (B) Southern blot analysis of wt and five T 2 transgenic lines (L1, L2, L3, L4, and L5), probed with hpt gene probe showing single copy insertion. (C) Semi quantitative RT-PCR showing overexpression of OsGS1;1 and OsGS2 in transgenic lines (L1, L4, and L5) as compared to wt . The rice eEF1α gene was used as a reference gene and rRNA was used as loading control. (D ; top panel) Immunoblot analysis of three transgenic rice lines (L1, L4, and L5) and wt using a recombinant antibody which detects both OsGS1;1 and OsGS2 isoforms (black lines separate spliced regions from same blot) ( D ; bottom panel) Coomassie blue stained Rubisco large subunit (RubL) was used as loading control. (E) Total GS activity of three transgenic rice lines (L1, L4, and L5) in comparison to wt ). One unit of GS activity represents 1.0 μmol of γ-glutamylhydroxamate produced in 20 min. Asterisks above bars indicate significant differences from wt (* at p ≤ 0.05 and ** at p ≤ 0.01).

    Article Snippet: The transformed calli were selected on Chu-N6 medium supplemented with 50 mg/L Hygromycin (Invitrogen, USA) and 1 mg/L PPT (Duchefa, Germany).

    Techniques: Transgenic Assay, Polymerase Chain Reaction, Amplification, Southern Blot, Quantitative RT-PCR, Over Expression, Recombinant, Staining, Activity Assay, Produced

    a Schematic diagram of FRT-GFP-globin constructs that lack or contain the human TNF 3′-UTR. Specified portions (573 nt) of the human TNF 3′-UTR were cloned downstream of the Globin-GFP-Globin reporter. b Site-directed recombination of the GFP-TNF-3′-UTR fusion gene into a single locus. Plasmids were cotransfected with a Flp recombinase expression vector (pOG44) into the Zeocin-resistant parental Flp-In-293 cell line (Parent FRT, Invitrogen) that contains a single FRT (Flp recombination target) site. The integration site is transcriptionally competent, with the GFP-TNF-3′-UTR fusion conferring hygromycin resistance and tetracycline inducibility

    Journal: Molecular biotechnology

    Article Title: A Flexible Approach to Studying Post-Transcriptional Gene Regulation in Stably Transfected Mammalian Cells

    doi: 10.1007/s12033-010-9360-8

    Figure Lengend Snippet: a Schematic diagram of FRT-GFP-globin constructs that lack or contain the human TNF 3′-UTR. Specified portions (573 nt) of the human TNF 3′-UTR were cloned downstream of the Globin-GFP-Globin reporter. b Site-directed recombination of the GFP-TNF-3′-UTR fusion gene into a single locus. Plasmids were cotransfected with a Flp recombinase expression vector (pOG44) into the Zeocin-resistant parental Flp-In-293 cell line (Parent FRT, Invitrogen) that contains a single FRT (Flp recombination target) site. The integration site is transcriptionally competent, with the GFP-TNF-3′-UTR fusion conferring hygromycin resistance and tetracycline inducibility

    Article Snippet: Cell lines containing stable integrants of the FRT-GFP-globin, FRT-GFP-TNF, FRT-GFP-ΔARE plasmids were generated by co-transfection with pOG44 (encoding the Flp recombinase) into parent FRT cells (Flp-In-T-REx-293 cells, Invitrogen) followed by hygromycin selection as described above.

    Techniques: Construct, Clone Assay, Expressing, Plasmid Preparation

    Mycobacterium tuberculosis cultured on 7H10 solid medium in the absence of the inducer ( Section 1 and Section 3 ) and the presence of 0.1 µg/mL pristinamycin ( Section 2 and Section 4 ). Plate B contained 100 µg/mL hygromycin. ( A ) MTB H37Rv bacteria formed a lawn of colonies on the medium and were dependent on the inducer for survival; ( B ) MTB:: asadh bacteria formed colonies only on the medium containing pristinamycin. The experiments were repeated three times, and typical images are shown.

    Journal: International Journal of Molecular Sciences

    Article Title: Identification and Validation of Aspartic Acid Semialdehyde Dehydrogenase as a New Anti-Mycobacterium Tuberculosis Target

    doi: 10.3390/ijms161023572

    Figure Lengend Snippet: Mycobacterium tuberculosis cultured on 7H10 solid medium in the absence of the inducer ( Section 1 and Section 3 ) and the presence of 0.1 µg/mL pristinamycin ( Section 2 and Section 4 ). Plate B contained 100 µg/mL hygromycin. ( A ) MTB H37Rv bacteria formed a lawn of colonies on the medium and were dependent on the inducer for survival; ( B ) MTB:: asadh bacteria formed colonies only on the medium containing pristinamycin. The experiments were repeated three times, and typical images are shown.

    Article Snippet: Hygromycin (Amresco, Anachem, Bedfordshire, UK) was added if necessary at 200 µg/mL for Escherichia coli and 100 µg/mL for Mycobacterium .

    Techniques: Cell Culture

    Genotypic and phenotypic analysis of genetically manipulated parasites. (A, B, and C) Genomic DNA from the following parasite strains was used as the template: lanes 1, wild type; lanes 2, ARG/arg heterozygote; lanes 3, Δ arg mutant 1; lanes 4, Δ arg mutant 2; lanes 5, Δ arg [ ARG ] mutant; lanes 6, Δ arg [ arg ΔAKL] mutant. (A) Primers designed to encompass the coding region were used to amplify the ARG gene. (B) The forward primer sequence was located in a region upstream of the 5′ flanking sequence (upstream of the targeting construct), and the reverse primer sequence was located within the hygromycin resistance gene. (C) The same sense primer as that used for panel B was used, and the antisense primer sequence was located within the phleomycin resistance gene. (D) Western blot analyses were performed with cell extracts prepared from wild-type, ARG/arg heterozygote, Δ arg mutant 1, Δ arg mutant 2, Δ arg [ ARG ], and Δ arg [ arg ΔAKL] parasites. Parasites were fractioned by SDS-PAGE and the blot probed with polyclonal antibodies against L. mexicana ARG and a commercial monoclonal antibody that recognizes tubulin. The tubulin antibody was employed to verify equal loading of protein on all lanes.

    Journal: Infection and Immunity

    Article Title: Arginase Is Essential for Survival of Leishmania donovani Promastigotes but Not Intracellular Amastigotes

    doi: 10.1128/IAI.00554-16

    Figure Lengend Snippet: Genotypic and phenotypic analysis of genetically manipulated parasites. (A, B, and C) Genomic DNA from the following parasite strains was used as the template: lanes 1, wild type; lanes 2, ARG/arg heterozygote; lanes 3, Δ arg mutant 1; lanes 4, Δ arg mutant 2; lanes 5, Δ arg [ ARG ] mutant; lanes 6, Δ arg [ arg ΔAKL] mutant. (A) Primers designed to encompass the coding region were used to amplify the ARG gene. (B) The forward primer sequence was located in a region upstream of the 5′ flanking sequence (upstream of the targeting construct), and the reverse primer sequence was located within the hygromycin resistance gene. (C) The same sense primer as that used for panel B was used, and the antisense primer sequence was located within the phleomycin resistance gene. (D) Western blot analyses were performed with cell extracts prepared from wild-type, ARG/arg heterozygote, Δ arg mutant 1, Δ arg mutant 2, Δ arg [ ARG ], and Δ arg [ arg ΔAKL] parasites. Parasites were fractioned by SDS-PAGE and the blot probed with polyclonal antibodies against L. mexicana ARG and a commercial monoclonal antibody that recognizes tubulin. The tubulin antibody was employed to verify equal loading of protein on all lanes.

    Article Snippet: Resazurin, G418, hygromycin, putrescine, and ornithine were purchased from VWR International (Radnor, PA).

    Techniques: Mutagenesis, Sequencing, Construct, Western Blot, SDS Page

    Southern blot analysis of integrated HIV-1 DNA demonstrates intact viral genomes. The integrated proviral HIV-1 DNA is depicted with the LTRs flanking the gag , pol , and hygromycin ( hygro ) genes. The numbers 1 and 9718 indicate the terminal nucleotide residues of the HXB2 genome. Numbers below the restriction enzymes ( Kas I, Xho I, Eco RV, Cla I, Bam HI, and Hin dIII) indicate the nucleotide positions of the respective cleavage sites in the HXB2 genome. Probes are indicated by a boxed region and correspond to a 534-bp sequence from the middle of the hygro gene (A) and HXB2 sequence from positions 641 to 828 (left end) and 8604 to 8895 (right end) (B). Horizontal arrows and their associated numbers indicated the expected sizes of restriction fragments for an intact proviral genome. The Southern blot analysis proceeded in two steps, first using a hygromycin probe to detect an intact genome between the two LTRs (A). The second step used individual probes for each end (B) to check for intact sequence to within 112 bp of the left end and to within 103 bp of the right end.

    Journal: Journal of Virology

    Article Title: Mutations in the Human Immunodeficiency Virus Type 1 Integrase D,D(35)E Motif Do Not Eliminate Provirus Formation

    doi:

    Figure Lengend Snippet: Southern blot analysis of integrated HIV-1 DNA demonstrates intact viral genomes. The integrated proviral HIV-1 DNA is depicted with the LTRs flanking the gag , pol , and hygromycin ( hygro ) genes. The numbers 1 and 9718 indicate the terminal nucleotide residues of the HXB2 genome. Numbers below the restriction enzymes ( Kas I, Xho I, Eco RV, Cla I, Bam HI, and Hin dIII) indicate the nucleotide positions of the respective cleavage sites in the HXB2 genome. Probes are indicated by a boxed region and correspond to a 534-bp sequence from the middle of the hygro gene (A) and HXB2 sequence from positions 641 to 828 (left end) and 8604 to 8895 (right end) (B). Horizontal arrows and their associated numbers indicated the expected sizes of restriction fragments for an intact proviral genome. The Southern blot analysis proceeded in two steps, first using a hygromycin probe to detect an intact genome between the two LTRs (A). The second step used individual probes for each end (B) to check for intact sequence to within 112 bp of the left end and to within 103 bp of the right end.

    Article Snippet: Clones of infected cells were generated by selection in complete medium containing 200 μg of hygromycin (Boehringer Mannheim) per ml, isolated by using cloning cylinders, and expanded in hygromycin selection.

    Techniques: Southern Blot, Sequencing

    Enhancement of vitamin B 6 content in rice by heterologous expression of Arabidopsis PDX 1/ PDX 2. (a) Schematic representation of the T‐ DNA regions of the transformation vector used to generate the 35S lines. LB , Left Border; RB , Right Border; 35S , cauliflower mosaic virus 35S ( Ca MV 35S ) promoter; At PDX 1.1 , At2g38230 coding sequence; At PDX2 , At5g60540 coding sequence; hpt II , hygromycin phosphotransferase gene; T1, Ca MV 35S terminator; T2, Octopine synthase terminator. (b) Vitamin B 6 content in leaf sample extracts of 35S lines in the T 2 generation compared to wild‐type ( TP 309) and empty vector control (p1300). Average ± SD of four biological replicates. Student's t‐ test (p1300 versus transgenic lines and TP 309), * P

    Journal: The Plant Journal

    Article Title: Enhancement of vitamin B6 levels in rice expressing Arabidopsis vitamin B6 biosynthesis de novo genes

    doi: 10.1111/tpj.14379

    Figure Lengend Snippet: Enhancement of vitamin B 6 content in rice by heterologous expression of Arabidopsis PDX 1/ PDX 2. (a) Schematic representation of the T‐ DNA regions of the transformation vector used to generate the 35S lines. LB , Left Border; RB , Right Border; 35S , cauliflower mosaic virus 35S ( Ca MV 35S ) promoter; At PDX 1.1 , At2g38230 coding sequence; At PDX2 , At5g60540 coding sequence; hpt II , hygromycin phosphotransferase gene; T1, Ca MV 35S terminator; T2, Octopine synthase terminator. (b) Vitamin B 6 content in leaf sample extracts of 35S lines in the T 2 generation compared to wild‐type ( TP 309) and empty vector control (p1300). Average ± SD of four biological replicates. Student's t‐ test (p1300 versus transgenic lines and TP 309), * P

    Article Snippet: Surface‐sterilized seeds were rinsed five times with sterile water before being placed in sterile plastic jars containing Murashige and Skoog (MS) medium (1× MS salts including vitamins (Duchefa, Haarlem, Netherlands), 3% (w/v) sucrose, 0.3% (w/v) gelrite; pH 5.8) containing 50 mg L−1 of hygromycin (Carl Roth, Karlsruhe, Germany) for selection of transgenic lines.

    Techniques: Expressing, Transformation Assay, Plasmid Preparation, Sequencing, Transgenic Assay

    EM sensitivity to the selective agents. Kan: kanamycin, Hyg: hygromycin. Data were collected after 3 subcultures (6 weeks) and the values for the 5 embryogenic lines were averaged.

    Journal: The Scientific World Journal

    Article Title: Stable Agrobacterium-Mediated Transformation of Maritime Pine Based on Kanamycin Selection

    doi: 10.1155/2013/681792

    Figure Lengend Snippet: EM sensitivity to the selective agents. Kan: kanamycin, Hyg: hygromycin. Data were collected after 3 subcultures (6 weeks) and the values for the 5 embryogenic lines were averaged.

    Article Snippet: Embryogenic Line Sensitivity to Selective Agents The selective agents kanamycin and hygromycin (Duchefa) were tested.

    Techniques:

    Structure and expression of the TAR1 gene. A, The TAR1 gene contains 14 exons (thick boxes) separated by 13 introns. The insertion site of the DNA cassette encoding the hygromycin resistance gene ( aphVII ) in tar1-1 is indicated by a black triangle. The

    Journal: Plant Physiology

    Article Title: Algal Dual-Specificity Tyrosine Phosphorylation-Regulated Kinase, Triacylglycerol Accumulation Regulator1, Regulates Accumulation of Triacylglycerol in Nitrogen or Sulfur Deficiency 1Algal Dual-Specificity Tyrosine Phosphorylation-Regulated Kinase, Triacylglycerol Accumulation Regulator1, Regulates Accumulation of Triacylglycerol in Nitrogen or Sulfur Deficiency 1 [OPEN]

    doi: 10.1104/pp.15.00319

    Figure Lengend Snippet: Structure and expression of the TAR1 gene. A, The TAR1 gene contains 14 exons (thick boxes) separated by 13 introns. The insertion site of the DNA cassette encoding the hygromycin resistance gene ( aphVII ) in tar1-1 is indicated by a black triangle. The

    Article Snippet: To prepare C. reinhardtii transformant pools for -mediated mutant screening, a 1,999-bp PCR fragment containing the hygromycin resistance gene aph7 expressed from the β2-tublin promoter was used to transform C-9 cells without cell wall removal as described previously ( ; ). solid medium containing 30 mg L−1 of hygromycin (Wako) was used for selection.

    Techniques: Expressing

    Modified Flp-In system. ( a ) A targeting vector is introduced to knock out a single allele of GSK-3β and create a host cell line with a FRT-puro cassette. Co-transfection of Flp recombinase and a gene of interest previously cloned into a vector containing a V5 epitope and FRT-hygro cassette results in homologous recombination at the FRT sites. Positive colonies are resistant to hygromycin and sensitive to puromycin. ( b ) Western blot analysis of transgenic myr-PDK-1 cell lines. ( c ) Membrane localization of myr-PDK-1 by immunofluorescent staining of V5; confocal, × 40 magnification.

    Journal: Oncogene

    Article Title: Activation of PDK-1 maintains mouse embryonic stem cell self-renewal in a PKB-dependent manner

    doi: 10.1038/onc.2013.44

    Figure Lengend Snippet: Modified Flp-In system. ( a ) A targeting vector is introduced to knock out a single allele of GSK-3β and create a host cell line with a FRT-puro cassette. Co-transfection of Flp recombinase and a gene of interest previously cloned into a vector containing a V5 epitope and FRT-hygro cassette results in homologous recombination at the FRT sites. Positive colonies are resistant to hygromycin and sensitive to puromycin. ( b ) Western blot analysis of transgenic myr-PDK-1 cell lines. ( c ) Membrane localization of myr-PDK-1 by immunofluorescent staining of V5; confocal, × 40 magnification.

    Article Snippet: Cells were placed in a six-well plate (Corning, Tewksbury, MA, USA) and selected with 250 μg/ml hygromycin (InvivoGen, San Diego, CA, USA) after 48 h. Colonies were picked and expanded from 96-well plates to replica 24-well plates, where 1.2 μg/ml puromycin selection medium was used in place of hygromycin for one of the replica plates.

    Techniques: Modification, Plasmid Preparation, Knock-Out, Cotransfection, Clone Assay, Homologous Recombination, Western Blot, Transgenic Assay, Staining

    Generation of the Δ msmHr M. smegmatis strain . (A) Genomic organization of the msmHr gene locus. (Upper panel) Genes are shown as large arrows in their native orientation. Small arrows represent the forward and reverse primers used for PCR, and sizes of the amplified products are indicated. Location and orientation of the hygromycin cassette are also indicated (Bottom panel). No PCR product was obtained using primers 2415InL and 2415InR to amplify the coding sequences of msmHr . PCR products for the upstream and downstream regions of msmHr were amplified using the primer pairs 2415LLL/IL(R) and 2415RRR/IR(F), respectively. (B) Effect of msmHr deletion on msmeg_2414 and msmeg_2416 expression. Quantitative real-time PCR (qRT-PCR) analysis of msmHr cluster transcription. The primer pairs 2414qF/2414qR and 2416qF/2416qR were used for qRT-PCR. Results are shown as the means ± standard deviations of three replicates ( * P

    Journal: Frontiers in Microbiology

    Article Title: A bacterial hemerythrin-like protein MsmHr inhibits the SigF-dependent hydrogen peroxide response in mycobacteria

    doi: 10.3389/fmicb.2014.00800

    Figure Lengend Snippet: Generation of the Δ msmHr M. smegmatis strain . (A) Genomic organization of the msmHr gene locus. (Upper panel) Genes are shown as large arrows in their native orientation. Small arrows represent the forward and reverse primers used for PCR, and sizes of the amplified products are indicated. Location and orientation of the hygromycin cassette are also indicated (Bottom panel). No PCR product was obtained using primers 2415InL and 2415InR to amplify the coding sequences of msmHr . PCR products for the upstream and downstream regions of msmHr were amplified using the primer pairs 2415LLL/IL(R) and 2415RRR/IR(F), respectively. (B) Effect of msmHr deletion on msmeg_2414 and msmeg_2416 expression. Quantitative real-time PCR (qRT-PCR) analysis of msmHr cluster transcription. The primer pairs 2414qF/2414qR and 2416qF/2416qR were used for qRT-PCR. Results are shown as the means ± standard deviations of three replicates ( * P

    Article Snippet: Hygromycin (75 mg/L for M. smegmatis , 150 mg/L for Escherichia coli ; Roche) and kanamycin (25 mg/L for M. smegmatis , 50 mg/L for Escherichia coli ; Amresco) were added to the medium as needed.

    Techniques: Polymerase Chain Reaction, Amplification, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Generation of a M. bovis BCG kasB null mutant. Maps of the kasB region in the M. bovis BCG genome and its corresponding region in the conditional mutant Δ kasB are shown on the left, with the corresponding Southern blot on the right. The regions used as porbes are indicated by solid lines with square ends while the expected bands are indicated by broken lines (with sizes indicated). hyg , hygromycin resistance gene from Streptomyces hygroscopicus ; res , γδ resolvase recognition sites.

    Journal: PLoS ONE

    Article Title: Platensimycin Activity against Mycobacterial ?-Ketoacyl-ACP Synthases

    doi: 10.1371/journal.pone.0006306

    Figure Lengend Snippet: Generation of a M. bovis BCG kasB null mutant. Maps of the kasB region in the M. bovis BCG genome and its corresponding region in the conditional mutant Δ kasB are shown on the left, with the corresponding Southern blot on the right. The regions used as porbes are indicated by solid lines with square ends while the expected bands are indicated by broken lines (with sizes indicated). hyg , hygromycin resistance gene from Streptomyces hygroscopicus ; res , γδ resolvase recognition sites.

    Article Snippet: The wells in rows B to G in columns 3 to 11 received 100 µl of 7H9 medium containing 25 µg/ml kanamycin, 50 µg/ml hygromycin and ADC (Beckton Dickinson, Sparks, MD).

    Techniques: Mutagenesis, Southern Blot

    Effects of the PDR1 YJM326 allele in different genetic backgrounds (A) Fitness variation of 20 isolates (left panel) in comparison with the same set of strains hybridized with YJM326 in the presence of drug. Fitness values (y-axis) correspond to the ratio between the growth in the presence of cycloheximide (YPD CHX 1µg/ml) and control media YPD. Dashed line indicates the fitness of the resistant strain YJM326. (B) Fitness variation of 20 isolates carrying empty control plasmid (pCTRL, left panel) or plasmid containing the PDR1 YJM326 allele under its native promoter (pPDR1 YJM326 , right panel). Fitness values were measured in the presence of cycloheximide (YPD CHX 1µg/ml) with hygromycin to maintain plasmid stability. Dashed line indicates the fitness value of YJM326 carrying the plasmid pPDR1 YJM326 for detailed comparison for effect of hybrid and plasmid in individual genetic backgrounds.

    Journal: Cell reports

    Article Title: The hidden complexity of Mendelian traits across yeast natural populations

    doi: 10.1016/j.celrep.2016.06.048

    Figure Lengend Snippet: Effects of the PDR1 YJM326 allele in different genetic backgrounds (A) Fitness variation of 20 isolates (left panel) in comparison with the same set of strains hybridized with YJM326 in the presence of drug. Fitness values (y-axis) correspond to the ratio between the growth in the presence of cycloheximide (YPD CHX 1µg/ml) and control media YPD. Dashed line indicates the fitness of the resistant strain YJM326. (B) Fitness variation of 20 isolates carrying empty control plasmid (pCTRL, left panel) or plasmid containing the PDR1 YJM326 allele under its native promoter (pPDR1 YJM326 , right panel). Fitness values were measured in the presence of cycloheximide (YPD CHX 1µg/ml) with hygromycin to maintain plasmid stability. Dashed line indicates the fitness value of YJM326 carrying the plasmid pPDR1 YJM326 for detailed comparison for effect of hybrid and plasmid in individual genetic backgrounds.

    Article Snippet: A final concentration of 200 µg/ml hygromycin (Euromedex) was supplemented to maintain the plasmids carrying a resistance marker gene HygMX .

    Techniques: Plasmid Preparation

    Generation of A . benhamiae Δ hapX mutants and reconstituted strains. (A) For deletion of the hapX locus (white arrow) in the wild-type strain A . benhamiae LAU2354-2 (bottom) a DNA cassette, containing the hygromycin resistance gene hph (black arrow) under control of the gpd promoter (P gpd , bent arrow) together with the termination sequence fragment T trpC (filled circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used (top). (B) For reinsertion of the hapX gene into its original locus in the Δ hapX mutants a DNA cassette, containing the coding region of hapX and the neomycin resistance gene neo (lined arrow) under control of the A . benhamiae actin promoter (P ACT1 , bent arrow) together with the Candida albicans actin termination sequence fragment T ACT1 (blank circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used. (C) Southern blot of Nde I-digested genomic DNA of the wild-type strain A . benhamiae LAU2354-2, Δ hapX mutants and hapX C reconstituted strains with hapX -specific probe 1. The probes which were used for Southern analysis of the transformants are indicated by the black bars. Only the following relevant restriction sites are given in panels a and b: A, Apa I; B, Bam HI; H, Hin dIII; Nd, Nde I; Not I. The sizes of the hybridizing fragments are given on the left and their identities on the right.

    Journal: PLoS ONE

    Article Title: HapX Mediates Iron Homeostasis in the Pathogenic Dermatophyte Arthroderma benhamiae but Is Dispensable for Virulence

    doi: 10.1371/journal.pone.0150701

    Figure Lengend Snippet: Generation of A . benhamiae Δ hapX mutants and reconstituted strains. (A) For deletion of the hapX locus (white arrow) in the wild-type strain A . benhamiae LAU2354-2 (bottom) a DNA cassette, containing the hygromycin resistance gene hph (black arrow) under control of the gpd promoter (P gpd , bent arrow) together with the termination sequence fragment T trpC (filled circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used (top). (B) For reinsertion of the hapX gene into its original locus in the Δ hapX mutants a DNA cassette, containing the coding region of hapX and the neomycin resistance gene neo (lined arrow) under control of the A . benhamiae actin promoter (P ACT1 , bent arrow) together with the Candida albicans actin termination sequence fragment T ACT1 (blank circle) flanked by hapX upstream and downstream regions ( hapX up and hapX down , solid lines), was used. (C) Southern blot of Nde I-digested genomic DNA of the wild-type strain A . benhamiae LAU2354-2, Δ hapX mutants and hapX C reconstituted strains with hapX -specific probe 1. The probes which were used for Southern analysis of the transformants are indicated by the black bars. Only the following relevant restriction sites are given in panels a and b: A, Apa I; B, Bam HI; H, Hin dIII; Nd, Nde I; Not I. The sizes of the hybridizing fragments are given on the left and their identities on the right.

    Article Snippet: For short-term storage, the wild-type A . benhamiae LAU2354-2 was cultivated on Sabouraud glucose agar [1% (w/v) peptone, 2% (w/v) glucose, 1.5% (w/v) agar] (SAB) and transformants of A . benhamiae LAU2354-2 were grown on SAB supplemented with 200 μg/ml hygromycin (ForMedium, Hunstanton, UK) or G418 (Carl Roth, Karlsruhe, Germany), according to the selectable marker used.

    Techniques: Sequencing, Southern Blot

    Manipulation of Abl expression and activity in normal and malignant MECs. A ) Immunoblotting for c-Abl expression in NMuMG and 4T1 cells engineered to express either a scrambled (scram) or an Abl-targeting shRNA (shAbl), or empty vector (pMSCV-hygromycin),

    Journal: The FASEB Journal

    Article Title: Activated Abl kinase inhibits oncogenic transforming growth factor-? signaling and tumorigenesis in mammary tumors

    doi: 10.1096/fj.09-138412

    Figure Lengend Snippet: Manipulation of Abl expression and activity in normal and malignant MECs. A ) Immunoblotting for c-Abl expression in NMuMG and 4T1 cells engineered to express either a scrambled (scram) or an Abl-targeting shRNA (shAbl), or empty vector (pMSCV-hygromycin),

    Article Snippet: Transduced cells were isolated by hygromycin selection (100 μg/ml; Mediatech, Manassas, VA, USA), and the resulting stable polyclonal populations of KD-Abl- or CST-Abl-expressing cells were expanded.

    Techniques: Expressing, Activity Assay, shRNA, Plasmid Preparation

    Establishment of core3 synthase-expressing HT-29 cells. A , HT-29 cells were transfected with mock and core3 synthase expression plasmids, and several hygromycin-resistant clones of each were isolated. A , RT-PCR results of core3 synthase expression of

    Journal: The Journal of Biological Chemistry

    Article Title: Core2 O-Glycan Structure Is Essential for the Cell Surface Expression of Sucrase Isomaltase and Dipeptidyl Peptidase-IV during Intestinal Cell Differentiation *

    doi: 10.1074/jbc.M110.162735

    Figure Lengend Snippet: Establishment of core3 synthase-expressing HT-29 cells. A , HT-29 cells were transfected with mock and core3 synthase expression plasmids, and several hygromycin-resistant clones of each were isolated. A , RT-PCR results of core3 synthase expression of

    Article Snippet: Selection was performed by a 2-week incubation in medium containing 1 mg/ml hygromycin (Promega).

    Techniques: Expressing, Transfection, Clone Assay, Isolation, Reverse Transcription Polymerase Chain Reaction