bsiwi  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Name:
    BsiWI
    Description:
    BsiWI 1 500 units
    Catalog Number:
    R0553L
    Price:
    282
    Category:
    Restriction Enzymes
    Size:
    1 500 units
    Buy from Supplier


    Structured Review

    New England Biolabs bsiwi
    BsiWI
    BsiWI 1 500 units
    https://www.bioz.com/result/bsiwi/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsiwi - by Bioz Stars, 2021-05
    94/100 stars

    Images

    1) Product Images from "Pathogenicity and Immunogenicity of Recombinant Rabies Viruses Expressing the Lagos Bat Virus Matrix and Glycoprotein: Perspectives for a Pan-Lyssavirus Vaccine"

    Article Title: Pathogenicity and Immunogenicity of Recombinant Rabies Viruses Expressing the Lagos Bat Virus Matrix and Glycoprotein: Perspectives for a Pan-Lyssavirus Vaccine

    Journal: Tropical Medicine and Infectious Disease

    doi: 10.3390/tropicalmed2030037

    Schematic representation of the construction of recombinant viruses. Restriction enzyme sites are indicated ( XmaI , PacI , AvrII , KpnI . BsiWI , AsiSI , NheI and AscI ). GAS represents the SPBN G gene with two amino acid substitutions (Asn 194 to Ser and Arg 333 to Glu). The following abbreviations were used: N, nucleoprotein; M, matrix protein; G, glycoprotein; L, RNA-dependent RNA polymerase. LBVM and LBVG represent the LBV M and G gene respectively.
    Figure Legend Snippet: Schematic representation of the construction of recombinant viruses. Restriction enzyme sites are indicated ( XmaI , PacI , AvrII , KpnI . BsiWI , AsiSI , NheI and AscI ). GAS represents the SPBN G gene with two amino acid substitutions (Asn 194 to Ser and Arg 333 to Glu). The following abbreviations were used: N, nucleoprotein; M, matrix protein; G, glycoprotein; L, RNA-dependent RNA polymerase. LBVM and LBVG represent the LBV M and G gene respectively.

    Techniques Used: Recombinant

    2) Product Images from "Overexpression of Tumor Necrosis Factor Alpha by a Recombinant Rabies Virus Attenuates Replication in Neurons and Prevents Lethal Infection in Mice"

    Article Title: Overexpression of Tumor Necrosis Factor Alpha by a Recombinant Rabies Virus Attenuates Replication in Neurons and Prevents Lethal Infection in Mice

    Journal: Journal of Virology

    doi: 10.1128/JVI.79.24.15405-15416.2005

    Expression of TNF-α by recombinant RV. (A) Schematic of RV-TNF-α recombinant constructs. The pSPBN vector was derived from SN by removing the Ψ gene and introducing BsiWI and NheI sites between the G and L genes. Mouse TNF-α cDNA was amplified by PCR, with the introduction of BsiWI and NheI sites, and was ligated into pSPBN, resulting in pSPBN-TNF-α(+). In pSPBN-TNF-α(MEM), the proteolytic cleavage site of TNF-α was removed by deleting the coding sequence for the first nine amino acids of soluble TNF-α and exchanging the lysine at position 11 with glutamic acid, as described in Materials and Methods. To construct pSPBN-TNF-α(−), the first seven ATG codons of the TNF-α gene were mutated. (B) TNF-α production in BSR cells. BSR cells were infected with SPBN-TNF-α(−), SPBN-TNF-α(MEM), or SPBN-TNF-α(+) at an MOI of 0.1 and incubated at 37°C. TNF-α secreted into the tissue culture supernatants of infected BSR cells was measured at 24 h p.i., using ELISA. OD, optical density. (C to L) Confocal double-immunofluorescence microscopy of TNF-α (green, panels C, F, and I) and rabies ribonucleocapsid (RNP) (red, panels D, G, and K) in NA cells infected with SPBN-TNF-α(+), SPBN-TNF-α(MEM), or SPBN-TNF-α(−). Panels E, H, and L show the composites of TNF-α and RNP double immunofluorescence. Note that TNF-α is absent only in NA cells infected with SPBN-TNF-α(−) (panels I and L).
    Figure Legend Snippet: Expression of TNF-α by recombinant RV. (A) Schematic of RV-TNF-α recombinant constructs. The pSPBN vector was derived from SN by removing the Ψ gene and introducing BsiWI and NheI sites between the G and L genes. Mouse TNF-α cDNA was amplified by PCR, with the introduction of BsiWI and NheI sites, and was ligated into pSPBN, resulting in pSPBN-TNF-α(+). In pSPBN-TNF-α(MEM), the proteolytic cleavage site of TNF-α was removed by deleting the coding sequence for the first nine amino acids of soluble TNF-α and exchanging the lysine at position 11 with glutamic acid, as described in Materials and Methods. To construct pSPBN-TNF-α(−), the first seven ATG codons of the TNF-α gene were mutated. (B) TNF-α production in BSR cells. BSR cells were infected with SPBN-TNF-α(−), SPBN-TNF-α(MEM), or SPBN-TNF-α(+) at an MOI of 0.1 and incubated at 37°C. TNF-α secreted into the tissue culture supernatants of infected BSR cells was measured at 24 h p.i., using ELISA. OD, optical density. (C to L) Confocal double-immunofluorescence microscopy of TNF-α (green, panels C, F, and I) and rabies ribonucleocapsid (RNP) (red, panels D, G, and K) in NA cells infected with SPBN-TNF-α(+), SPBN-TNF-α(MEM), or SPBN-TNF-α(−). Panels E, H, and L show the composites of TNF-α and RNP double immunofluorescence. Note that TNF-α is absent only in NA cells infected with SPBN-TNF-α(−) (panels I and L).

    Techniques Used: Expressing, Recombinant, Construct, Plasmid Preparation, Derivative Assay, Amplification, Polymerase Chain Reaction, Sequencing, Infection, Incubation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Microscopy

    3) Product Images from "The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial ▿"

    Article Title: The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.01346-09

    Construction and characterization of recombinant RABVs expressing different chemokins. (A) Construction of full-length recombinant RABVs. Chemokine genes MIP-1α, RANTES, and IP-10 were individually inserted between BsiWI and NheI sites of the
    Figure Legend Snippet: Construction and characterization of recombinant RABVs expressing different chemokins. (A) Construction of full-length recombinant RABVs. Chemokine genes MIP-1α, RANTES, and IP-10 were individually inserted between BsiWI and NheI sites of the

    Techniques Used: Recombinant, Expressing

    4) Product Images from "High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases"

    Article Title: High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1004090

    Herpes simplex viral genome editing targeted by the CRISPR-Cas system. (A) Schematic of the HSV1 genome and the gRNA-206 target site in the UL23 gene. PAM sequences are highlighted in orange. The BsiWI restriction site used for RFLP is highlighted in aqua green. (B) The fragment amplified from an HSV1 genome using oligo 8 and oligo 9 was used in the RFLP assay. Co-expression of Cas9 and gRNA-206 in HSV1-infected cells leads to indel mutations in the TK gene of the HSV1 genome, which destroys the BsiWI restriction site. Arrows indicate fragments generated by BsiWI digestion. (C) Titration of HSV1 using an endpoint dilution assay in the presence or absence of 100 µg/ml ACV. The titers of total HSV1 progeny are represented by a blue bar. The titers of ACV-resistant HSV1 progeny virus are represented by an orange bar. N = 3; error bars show the means ± SEM. ND, not detected. (D) Sequences of indel mutations identified from eight ACV-resistant HSV1 strains. Red dashes, deletions; red bases, insertions. The incidence of each genotype is listed in the right-most column. (E) Strategy of Cas9:gRNA-induced HDR used to insert the EGFP gene into the HSV1 genome using donor DNA. (F) Titration of recombinant HSV1 using a plaque assay. HDR was induced by the Cas9 protein or Cas9:gRNA206. Blue bars represent the total number of macroscopically visible plaques; and green bars represent the number of green fluorescent plaques that were counted using a fluorescence microscope. N > 5; error bars show the means ± SEM. ND, not detected. (G) An isolated green HSV1 plaque that was observed using a fluorescence microscope and merged with an image of DAPI staining. Scale bar, 50 µm. (H) PCR amplification of HSV1 genomes extracted from a green fluorescent plaque using oligo 8 and oligo 9 to verify the lack of wild-type HSV1 contamination.
    Figure Legend Snippet: Herpes simplex viral genome editing targeted by the CRISPR-Cas system. (A) Schematic of the HSV1 genome and the gRNA-206 target site in the UL23 gene. PAM sequences are highlighted in orange. The BsiWI restriction site used for RFLP is highlighted in aqua green. (B) The fragment amplified from an HSV1 genome using oligo 8 and oligo 9 was used in the RFLP assay. Co-expression of Cas9 and gRNA-206 in HSV1-infected cells leads to indel mutations in the TK gene of the HSV1 genome, which destroys the BsiWI restriction site. Arrows indicate fragments generated by BsiWI digestion. (C) Titration of HSV1 using an endpoint dilution assay in the presence or absence of 100 µg/ml ACV. The titers of total HSV1 progeny are represented by a blue bar. The titers of ACV-resistant HSV1 progeny virus are represented by an orange bar. N = 3; error bars show the means ± SEM. ND, not detected. (D) Sequences of indel mutations identified from eight ACV-resistant HSV1 strains. Red dashes, deletions; red bases, insertions. The incidence of each genotype is listed in the right-most column. (E) Strategy of Cas9:gRNA-induced HDR used to insert the EGFP gene into the HSV1 genome using donor DNA. (F) Titration of recombinant HSV1 using a plaque assay. HDR was induced by the Cas9 protein or Cas9:gRNA206. Blue bars represent the total number of macroscopically visible plaques; and green bars represent the number of green fluorescent plaques that were counted using a fluorescence microscope. N > 5; error bars show the means ± SEM. ND, not detected. (G) An isolated green HSV1 plaque that was observed using a fluorescence microscope and merged with an image of DAPI staining. Scale bar, 50 µm. (H) PCR amplification of HSV1 genomes extracted from a green fluorescent plaque using oligo 8 and oligo 9 to verify the lack of wild-type HSV1 contamination.

    Techniques Used: CRISPR, Amplification, RFLP Assay, Expressing, Infection, Generated, Titration, Endpoint Dilution Assay, Recombinant, Plaque Assay, Fluorescence, Microscopy, Isolation, Staining, Polymerase Chain Reaction

    5) Product Images from "Overexpression of Tumor Necrosis Factor Alpha by a Recombinant Rabies Virus Attenuates Replication in Neurons and Prevents Lethal Infection in Mice"

    Article Title: Overexpression of Tumor Necrosis Factor Alpha by a Recombinant Rabies Virus Attenuates Replication in Neurons and Prevents Lethal Infection in Mice

    Journal: Journal of Virology

    doi: 10.1128/JVI.79.24.15405-15416.2005

    Expression of TNF-α by recombinant RV. (A) Schematic of RV-TNF-α recombinant constructs. The pSPBN vector was derived from SN by removing the Ψ gene and introducing BsiWI and NheI sites between the G and L genes. Mouse TNF-α cDNA was amplified by PCR, with the introduction of BsiWI and NheI sites, and was ligated into pSPBN, resulting in pSPBN-TNF-α(+). In pSPBN-TNF-α(MEM), the proteolytic cleavage site of TNF-α was removed by deleting the coding sequence for the first nine amino acids of soluble TNF-α and exchanging the lysine at position 11 with glutamic acid, as described in Materials and Methods. To construct pSPBN-TNF-α(−), the first seven ATG codons of the TNF-α gene were mutated. (B) TNF-α production in BSR cells. BSR cells were infected with SPBN-TNF-α(−), SPBN-TNF-α(MEM), or SPBN-TNF-α(+) at an MOI of 0.1 and incubated at 37°C. TNF-α secreted into the tissue culture supernatants of infected BSR cells was measured at 24 h p.i., using ELISA. OD, optical density. (C to L) Confocal double-immunofluorescence microscopy of TNF-α (green, panels C, F, and I) and rabies ribonucleocapsid (RNP) (red, panels D, G, and K) in NA cells infected with SPBN-TNF-α(+), SPBN-TNF-α(MEM), or SPBN-TNF-α(−). Panels E, H, and L show the composites of TNF-α and RNP double immunofluorescence. Note that TNF-α is absent only in NA cells infected with SPBN-TNF-α(−) (panels I and L).
    Figure Legend Snippet: Expression of TNF-α by recombinant RV. (A) Schematic of RV-TNF-α recombinant constructs. The pSPBN vector was derived from SN by removing the Ψ gene and introducing BsiWI and NheI sites between the G and L genes. Mouse TNF-α cDNA was amplified by PCR, with the introduction of BsiWI and NheI sites, and was ligated into pSPBN, resulting in pSPBN-TNF-α(+). In pSPBN-TNF-α(MEM), the proteolytic cleavage site of TNF-α was removed by deleting the coding sequence for the first nine amino acids of soluble TNF-α and exchanging the lysine at position 11 with glutamic acid, as described in Materials and Methods. To construct pSPBN-TNF-α(−), the first seven ATG codons of the TNF-α gene were mutated. (B) TNF-α production in BSR cells. BSR cells were infected with SPBN-TNF-α(−), SPBN-TNF-α(MEM), or SPBN-TNF-α(+) at an MOI of 0.1 and incubated at 37°C. TNF-α secreted into the tissue culture supernatants of infected BSR cells was measured at 24 h p.i., using ELISA. OD, optical density. (C to L) Confocal double-immunofluorescence microscopy of TNF-α (green, panels C, F, and I) and rabies ribonucleocapsid (RNP) (red, panels D, G, and K) in NA cells infected with SPBN-TNF-α(+), SPBN-TNF-α(MEM), or SPBN-TNF-α(−). Panels E, H, and L show the composites of TNF-α and RNP double immunofluorescence. Note that TNF-α is absent only in NA cells infected with SPBN-TNF-α(−) (panels I and L).

    Techniques Used: Expressing, Recombinant, Construct, Plasmid Preparation, Derivative Assay, Amplification, Polymerase Chain Reaction, Sequencing, Infection, Incubation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Microscopy

    6) Product Images from "The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial ▿"

    Article Title: The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial ▿

    Journal: Journal of Virology

    doi: 10.1128/JVI.01346-09

    Construction and characterization of recombinant RABVs expressing different chemokins. (A) Construction of full-length recombinant RABVs. Chemokine genes MIP-1α, RANTES, and IP-10 were individually inserted between BsiWI and NheI sites of the
    Figure Legend Snippet: Construction and characterization of recombinant RABVs expressing different chemokins. (A) Construction of full-length recombinant RABVs. Chemokine genes MIP-1α, RANTES, and IP-10 were individually inserted between BsiWI and NheI sites of the

    Techniques Used: Recombinant, Expressing

    7) Product Images from "Pathogenicity and Immunogenicity of Recombinant Rabies Viruses Expressing the Lagos Bat Virus Matrix and Glycoprotein: Perspectives for a Pan-Lyssavirus Vaccine"

    Article Title: Pathogenicity and Immunogenicity of Recombinant Rabies Viruses Expressing the Lagos Bat Virus Matrix and Glycoprotein: Perspectives for a Pan-Lyssavirus Vaccine

    Journal: Tropical Medicine and Infectious Disease

    doi: 10.3390/tropicalmed2030037

    Schematic representation of the construction of recombinant viruses. Restriction enzyme sites are indicated ( XmaI , PacI , AvrII , KpnI . BsiWI , AsiSI , NheI and AscI ). GAS represents the SPBN G gene with two amino acid substitutions (Asn 194 to Ser and Arg 333 to Glu). The following abbreviations were used: N, nucleoprotein; M, matrix protein; G, glycoprotein; L, RNA-dependent RNA polymerase. LBVM and LBVG represent the LBV M and G gene respectively.
    Figure Legend Snippet: Schematic representation of the construction of recombinant viruses. Restriction enzyme sites are indicated ( XmaI , PacI , AvrII , KpnI . BsiWI , AsiSI , NheI and AscI ). GAS represents the SPBN G gene with two amino acid substitutions (Asn 194 to Ser and Arg 333 to Glu). The following abbreviations were used: N, nucleoprotein; M, matrix protein; G, glycoprotein; L, RNA-dependent RNA polymerase. LBVM and LBVG represent the LBV M and G gene respectively.

    Techniques Used: Recombinant

    Related Articles

    Polymerase Chain Reaction:

    Article Title: A Recombinant Rabies Virus Expressing Fms-like Tyrosine Kinase 3 Ligand (Flt3L) Induces Enhanced Immunogenicity in Mice
    Article Snippet: .. Briefly, PCR production ( Flt3L gene) and LBNSE infectious clone were both digested with Bsi WI and Nhe I (New England BioLabs, Ipswich, MA, USA), and after recycling, the Flt3L gene was inserted between G and L genes of LBNSE by ligation to finish the construction of LBNSE-Flt3L infectious clone. .. For the rescue of LBNSE-Flt3L, BSR cells were transfected with 2.0 µg of LBNSE-Flt3L infectious clone, 0.5 µg of pH-N, 0.25 µg of pH-P, 0.15 µg of pH-G, and 0.1 µg of pH-L using the SuperFect transfection reagent (Qiagen) according to the manufacturer’s protocol.

    Article Title: The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial ▿
    Article Snippet: The primer sets used for PCR were designed by Primer3 ( ) (Table ). .. The PCR products were digested with BsiWI and NheI (New England Biolabs, Berverly, MA) and then ligated into RABV vector pHEP-3.0 ( ) that had been previously digested with BsiWI and NheI. .. The resulting plasmids had each of the chemokine genes cloned between RABV glycoprotein (G) and the polymerase (L) genes and were designated pHEP-MIP1α, pHEP-RANTES, and pHEP-IP10, respectively (Fig. ).

    Article Title: Overexpression of Tumor Necrosis Factor Alpha by a Recombinant Rabies Virus Attenuates Replication in Neurons and Prevents Lethal Infection in Mice
    Article Snippet: Mouse TNF-α cDNA was amplified using Expand Hi-Fi Taq polymerase (Roche Applied Science, Indianapolis, IN) and the gene-specific primers TNF-1(+) (5′-AAA CGTACG ATG AGCACAGAAAGCATGATC-3′ [a BsiWI site is underlined, and the start codon is shown in bold]) and TNF-2(−) (5′-AAA GCTAGC TCA CAGAGCAATGACTCCAAAG-3′ [an NheI site is underlined, and the stop codon is shown in bold]) to introduce BsiWI and NheI recognition sites before and after the TNF-α coding region. .. The PCR product was digested with BsiWI and NheI (New England Biolabs, Beverly, MA) and ligated into RV vector pSPBN that had been previously digested with BsiWI and NheI. .. The resulting plasmid was designated pSPBN-TNF-α(+) (Fig. ).

    Article Title: High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases
    Article Snippet: The PCR products were ligated into the pMD20-T vector (Takara Bio Inc.) and submitted for sequencing using universal primers (BGI, Guangzhou, China). .. Restriction fragment length polymorphism (RFLP) analysis of gene modifications Purified HSV1 DNA products from the genomic DNA extraction and PCR amplification were digested with BsiWI (New England Biolabs) for 16 hours at 55°C and analyzed on 0.5 µg/ml ethidium bromide-stained agarose gels (1%). ..

    Article Title: Overexpression of Cytochrome c by a Recombinant Rabies Virus Attenuates Pathogenicity and Enhances Antiviral Immunity
    Article Snippet: Human cytochrome c cDNA was amplified using Eppendorf Taq DNA polymerase (Fisher Scientific, Pittsburgh, Pa.) and primers Cyt 5 (5′-AAA CGTACG AATATGGGTGATGTTGAGAA-3′ [ Bsi WI site underlined]) and Cyt 3 (5′-GAA GCTAGC TTACTCATTAGTAGCTTTTTTGAG-3′ [ Nhe I site underlined]) to introduce Bsi WI and Nhe I recognition sites before and after the cytochrome c coding region. .. The PCR product was digested with Bsi WI and Nhe I (New England Biolabs) and ligated into rabies virus vector pSPBN, which had been digested with Bsi WI and Nhe I ( ). .. The resulting plasmid was designated pSPBN-Cyto c (+) (Fig. ).

    Article Title: Pathogenicity and Immunogenicity of Recombinant Rabies Viruses Expressing the Lagos Bat Virus Matrix and Glycoprotein: Perspectives for a Pan-Lyssavirus Vaccine
    Article Snippet: To construct the recombinant virus with a LBVAFR1999 G gene sandwiched between two RABV G genes, the G gene was amplified from the LBVAFR1999 RNA using Pfu DNA polymerase (Promega, Madison, WI, USA) and the primers LBVEcoRIBsiWI and LBVXbaIAsiSI ( ). .. The amplified PCR product was digested with BsiWI and AsiSI (New England Biolabs, Ipswich, MA, USA) and then ligated to pSPBNGAS-GAS-GAS previously digested with BsiWI and AsiSI . .. The resulting plasmid was designated pSPBNGAS-LBVG-GAS ( ).

    Ligation:

    Article Title: A Recombinant Rabies Virus Expressing Fms-like Tyrosine Kinase 3 Ligand (Flt3L) Induces Enhanced Immunogenicity in Mice
    Article Snippet: .. Briefly, PCR production ( Flt3L gene) and LBNSE infectious clone were both digested with Bsi WI and Nhe I (New England BioLabs, Ipswich, MA, USA), and after recycling, the Flt3L gene was inserted between G and L genes of LBNSE by ligation to finish the construction of LBNSE-Flt3L infectious clone. .. For the rescue of LBNSE-Flt3L, BSR cells were transfected with 2.0 µg of LBNSE-Flt3L infectious clone, 0.5 µg of pH-N, 0.25 µg of pH-P, 0.15 µg of pH-G, and 0.1 µg of pH-L using the SuperFect transfection reagent (Qiagen) according to the manufacturer’s protocol.

    Plasmid Preparation:

    Article Title: The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial ▿
    Article Snippet: The primer sets used for PCR were designed by Primer3 ( ) (Table ). .. The PCR products were digested with BsiWI and NheI (New England Biolabs, Berverly, MA) and then ligated into RABV vector pHEP-3.0 ( ) that had been previously digested with BsiWI and NheI. .. The resulting plasmids had each of the chemokine genes cloned between RABV glycoprotein (G) and the polymerase (L) genes and were designated pHEP-MIP1α, pHEP-RANTES, and pHEP-IP10, respectively (Fig. ).

    Article Title: Overexpression of Tumor Necrosis Factor Alpha by a Recombinant Rabies Virus Attenuates Replication in Neurons and Prevents Lethal Infection in Mice
    Article Snippet: Mouse TNF-α cDNA was amplified using Expand Hi-Fi Taq polymerase (Roche Applied Science, Indianapolis, IN) and the gene-specific primers TNF-1(+) (5′-AAA CGTACG ATG AGCACAGAAAGCATGATC-3′ [a BsiWI site is underlined, and the start codon is shown in bold]) and TNF-2(−) (5′-AAA GCTAGC TCA CAGAGCAATGACTCCAAAG-3′ [an NheI site is underlined, and the stop codon is shown in bold]) to introduce BsiWI and NheI recognition sites before and after the TNF-α coding region. .. The PCR product was digested with BsiWI and NheI (New England Biolabs, Beverly, MA) and ligated into RV vector pSPBN that had been previously digested with BsiWI and NheI. .. The resulting plasmid was designated pSPBN-TNF-α(+) (Fig. ).

    Article Title: Overexpression of Cytochrome c by a Recombinant Rabies Virus Attenuates Pathogenicity and Enhances Antiviral Immunity
    Article Snippet: Human cytochrome c cDNA was amplified using Eppendorf Taq DNA polymerase (Fisher Scientific, Pittsburgh, Pa.) and primers Cyt 5 (5′-AAA CGTACG AATATGGGTGATGTTGAGAA-3′ [ Bsi WI site underlined]) and Cyt 3 (5′-GAA GCTAGC TTACTCATTAGTAGCTTTTTTGAG-3′ [ Nhe I site underlined]) to introduce Bsi WI and Nhe I recognition sites before and after the cytochrome c coding region. .. The PCR product was digested with Bsi WI and Nhe I (New England Biolabs) and ligated into rabies virus vector pSPBN, which had been digested with Bsi WI and Nhe I ( ). .. The resulting plasmid was designated pSPBN-Cyto c (+) (Fig. ).

    Purification:

    Article Title: High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases
    Article Snippet: The PCR products were ligated into the pMD20-T vector (Takara Bio Inc.) and submitted for sequencing using universal primers (BGI, Guangzhou, China). .. Restriction fragment length polymorphism (RFLP) analysis of gene modifications Purified HSV1 DNA products from the genomic DNA extraction and PCR amplification were digested with BsiWI (New England Biolabs) for 16 hours at 55°C and analyzed on 0.5 µg/ml ethidium bromide-stained agarose gels (1%). ..

    DNA Extraction:

    Article Title: High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases
    Article Snippet: The PCR products were ligated into the pMD20-T vector (Takara Bio Inc.) and submitted for sequencing using universal primers (BGI, Guangzhou, China). .. Restriction fragment length polymorphism (RFLP) analysis of gene modifications Purified HSV1 DNA products from the genomic DNA extraction and PCR amplification were digested with BsiWI (New England Biolabs) for 16 hours at 55°C and analyzed on 0.5 µg/ml ethidium bromide-stained agarose gels (1%). ..

    Amplification:

    Article Title: High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases
    Article Snippet: The PCR products were ligated into the pMD20-T vector (Takara Bio Inc.) and submitted for sequencing using universal primers (BGI, Guangzhou, China). .. Restriction fragment length polymorphism (RFLP) analysis of gene modifications Purified HSV1 DNA products from the genomic DNA extraction and PCR amplification were digested with BsiWI (New England Biolabs) for 16 hours at 55°C and analyzed on 0.5 µg/ml ethidium bromide-stained agarose gels (1%). ..

    Article Title: Pathogenicity and Immunogenicity of Recombinant Rabies Viruses Expressing the Lagos Bat Virus Matrix and Glycoprotein: Perspectives for a Pan-Lyssavirus Vaccine
    Article Snippet: To construct the recombinant virus with a LBVAFR1999 G gene sandwiched between two RABV G genes, the G gene was amplified from the LBVAFR1999 RNA using Pfu DNA polymerase (Promega, Madison, WI, USA) and the primers LBVEcoRIBsiWI and LBVXbaIAsiSI ( ). .. The amplified PCR product was digested with BsiWI and AsiSI (New England Biolabs, Ipswich, MA, USA) and then ligated to pSPBNGAS-GAS-GAS previously digested with BsiWI and AsiSI . .. The resulting plasmid was designated pSPBNGAS-LBVG-GAS ( ).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    New England Biolabs bsiwi
    Schematic representation of the construction of recombinant viruses. Restriction enzyme sites are indicated ( XmaI , PacI , AvrII , KpnI . <t>BsiWI</t> , <t>AsiSI</t> , NheI and AscI ). GAS represents the SPBN G gene with two amino acid substitutions (Asn 194 to Ser and Arg 333 to Glu). The following abbreviations were used: N, nucleoprotein; M, matrix protein; G, glycoprotein; L, RNA-dependent RNA polymerase. LBVM and LBVG represent the LBV M and G gene respectively.
    Bsiwi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsiwi/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsiwi - by Bioz Stars, 2021-05
    94/100 stars
      Buy from Supplier

    Image Search Results


    Schematic representation of the construction of recombinant viruses. Restriction enzyme sites are indicated ( XmaI , PacI , AvrII , KpnI . BsiWI , AsiSI , NheI and AscI ). GAS represents the SPBN G gene with two amino acid substitutions (Asn 194 to Ser and Arg 333 to Glu). The following abbreviations were used: N, nucleoprotein; M, matrix protein; G, glycoprotein; L, RNA-dependent RNA polymerase. LBVM and LBVG represent the LBV M and G gene respectively.

    Journal: Tropical Medicine and Infectious Disease

    Article Title: Pathogenicity and Immunogenicity of Recombinant Rabies Viruses Expressing the Lagos Bat Virus Matrix and Glycoprotein: Perspectives for a Pan-Lyssavirus Vaccine

    doi: 10.3390/tropicalmed2030037

    Figure Lengend Snippet: Schematic representation of the construction of recombinant viruses. Restriction enzyme sites are indicated ( XmaI , PacI , AvrII , KpnI . BsiWI , AsiSI , NheI and AscI ). GAS represents the SPBN G gene with two amino acid substitutions (Asn 194 to Ser and Arg 333 to Glu). The following abbreviations were used: N, nucleoprotein; M, matrix protein; G, glycoprotein; L, RNA-dependent RNA polymerase. LBVM and LBVG represent the LBV M and G gene respectively.

    Article Snippet: The amplified PCR product was digested with BsiWI and AsiSI (New England Biolabs, Ipswich, MA, USA) and then ligated to pSPBNGAS-GAS-GAS previously digested with BsiWI and AsiSI .

    Techniques: Recombinant

    Expression of TNF-α by recombinant RV. (A) Schematic of RV-TNF-α recombinant constructs. The pSPBN vector was derived from SN by removing the Ψ gene and introducing BsiWI and NheI sites between the G and L genes. Mouse TNF-α cDNA was amplified by PCR, with the introduction of BsiWI and NheI sites, and was ligated into pSPBN, resulting in pSPBN-TNF-α(+). In pSPBN-TNF-α(MEM), the proteolytic cleavage site of TNF-α was removed by deleting the coding sequence for the first nine amino acids of soluble TNF-α and exchanging the lysine at position 11 with glutamic acid, as described in Materials and Methods. To construct pSPBN-TNF-α(−), the first seven ATG codons of the TNF-α gene were mutated. (B) TNF-α production in BSR cells. BSR cells were infected with SPBN-TNF-α(−), SPBN-TNF-α(MEM), or SPBN-TNF-α(+) at an MOI of 0.1 and incubated at 37°C. TNF-α secreted into the tissue culture supernatants of infected BSR cells was measured at 24 h p.i., using ELISA. OD, optical density. (C to L) Confocal double-immunofluorescence microscopy of TNF-α (green, panels C, F, and I) and rabies ribonucleocapsid (RNP) (red, panels D, G, and K) in NA cells infected with SPBN-TNF-α(+), SPBN-TNF-α(MEM), or SPBN-TNF-α(−). Panels E, H, and L show the composites of TNF-α and RNP double immunofluorescence. Note that TNF-α is absent only in NA cells infected with SPBN-TNF-α(−) (panels I and L).

    Journal: Journal of Virology

    Article Title: Overexpression of Tumor Necrosis Factor Alpha by a Recombinant Rabies Virus Attenuates Replication in Neurons and Prevents Lethal Infection in Mice

    doi: 10.1128/JVI.79.24.15405-15416.2005

    Figure Lengend Snippet: Expression of TNF-α by recombinant RV. (A) Schematic of RV-TNF-α recombinant constructs. The pSPBN vector was derived from SN by removing the Ψ gene and introducing BsiWI and NheI sites between the G and L genes. Mouse TNF-α cDNA was amplified by PCR, with the introduction of BsiWI and NheI sites, and was ligated into pSPBN, resulting in pSPBN-TNF-α(+). In pSPBN-TNF-α(MEM), the proteolytic cleavage site of TNF-α was removed by deleting the coding sequence for the first nine amino acids of soluble TNF-α and exchanging the lysine at position 11 with glutamic acid, as described in Materials and Methods. To construct pSPBN-TNF-α(−), the first seven ATG codons of the TNF-α gene were mutated. (B) TNF-α production in BSR cells. BSR cells were infected with SPBN-TNF-α(−), SPBN-TNF-α(MEM), or SPBN-TNF-α(+) at an MOI of 0.1 and incubated at 37°C. TNF-α secreted into the tissue culture supernatants of infected BSR cells was measured at 24 h p.i., using ELISA. OD, optical density. (C to L) Confocal double-immunofluorescence microscopy of TNF-α (green, panels C, F, and I) and rabies ribonucleocapsid (RNP) (red, panels D, G, and K) in NA cells infected with SPBN-TNF-α(+), SPBN-TNF-α(MEM), or SPBN-TNF-α(−). Panels E, H, and L show the composites of TNF-α and RNP double immunofluorescence. Note that TNF-α is absent only in NA cells infected with SPBN-TNF-α(−) (panels I and L).

    Article Snippet: The PCR product was digested with BsiWI and NheI (New England Biolabs, Beverly, MA) and ligated into RV vector pSPBN that had been previously digested with BsiWI and NheI.

    Techniques: Expressing, Recombinant, Construct, Plasmid Preparation, Derivative Assay, Amplification, Polymerase Chain Reaction, Sequencing, Infection, Incubation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Microscopy

    Construction and characterization of recombinant RABVs expressing different chemokins. (A) Construction of full-length recombinant RABVs. Chemokine genes MIP-1α, RANTES, and IP-10 were individually inserted between BsiWI and NheI sites of the

    Journal: Journal of Virology

    Article Title: The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial ▿

    doi: 10.1128/JVI.01346-09

    Figure Lengend Snippet: Construction and characterization of recombinant RABVs expressing different chemokins. (A) Construction of full-length recombinant RABVs. Chemokine genes MIP-1α, RANTES, and IP-10 were individually inserted between BsiWI and NheI sites of the

    Article Snippet: The PCR products were digested with BsiWI and NheI (New England Biolabs, Berverly, MA) and then ligated into RABV vector pHEP-3.0 ( ) that had been previously digested with BsiWI and NheI.

    Techniques: Recombinant, Expressing

    Herpes simplex viral genome editing targeted by the CRISPR-Cas system. (A) Schematic of the HSV1 genome and the gRNA-206 target site in the UL23 gene. PAM sequences are highlighted in orange. The BsiWI restriction site used for RFLP is highlighted in aqua green. (B) The fragment amplified from an HSV1 genome using oligo 8 and oligo 9 was used in the RFLP assay. Co-expression of Cas9 and gRNA-206 in HSV1-infected cells leads to indel mutations in the TK gene of the HSV1 genome, which destroys the BsiWI restriction site. Arrows indicate fragments generated by BsiWI digestion. (C) Titration of HSV1 using an endpoint dilution assay in the presence or absence of 100 µg/ml ACV. The titers of total HSV1 progeny are represented by a blue bar. The titers of ACV-resistant HSV1 progeny virus are represented by an orange bar. N = 3; error bars show the means ± SEM. ND, not detected. (D) Sequences of indel mutations identified from eight ACV-resistant HSV1 strains. Red dashes, deletions; red bases, insertions. The incidence of each genotype is listed in the right-most column. (E) Strategy of Cas9:gRNA-induced HDR used to insert the EGFP gene into the HSV1 genome using donor DNA. (F) Titration of recombinant HSV1 using a plaque assay. HDR was induced by the Cas9 protein or Cas9:gRNA206. Blue bars represent the total number of macroscopically visible plaques; and green bars represent the number of green fluorescent plaques that were counted using a fluorescence microscope. N > 5; error bars show the means ± SEM. ND, not detected. (G) An isolated green HSV1 plaque that was observed using a fluorescence microscope and merged with an image of DAPI staining. Scale bar, 50 µm. (H) PCR amplification of HSV1 genomes extracted from a green fluorescent plaque using oligo 8 and oligo 9 to verify the lack of wild-type HSV1 contamination.

    Journal: PLoS Pathogens

    Article Title: High-Efficiency Targeted Editing of Large Viral Genomes by RNA-Guided Nucleases

    doi: 10.1371/journal.ppat.1004090

    Figure Lengend Snippet: Herpes simplex viral genome editing targeted by the CRISPR-Cas system. (A) Schematic of the HSV1 genome and the gRNA-206 target site in the UL23 gene. PAM sequences are highlighted in orange. The BsiWI restriction site used for RFLP is highlighted in aqua green. (B) The fragment amplified from an HSV1 genome using oligo 8 and oligo 9 was used in the RFLP assay. Co-expression of Cas9 and gRNA-206 in HSV1-infected cells leads to indel mutations in the TK gene of the HSV1 genome, which destroys the BsiWI restriction site. Arrows indicate fragments generated by BsiWI digestion. (C) Titration of HSV1 using an endpoint dilution assay in the presence or absence of 100 µg/ml ACV. The titers of total HSV1 progeny are represented by a blue bar. The titers of ACV-resistant HSV1 progeny virus are represented by an orange bar. N = 3; error bars show the means ± SEM. ND, not detected. (D) Sequences of indel mutations identified from eight ACV-resistant HSV1 strains. Red dashes, deletions; red bases, insertions. The incidence of each genotype is listed in the right-most column. (E) Strategy of Cas9:gRNA-induced HDR used to insert the EGFP gene into the HSV1 genome using donor DNA. (F) Titration of recombinant HSV1 using a plaque assay. HDR was induced by the Cas9 protein or Cas9:gRNA206. Blue bars represent the total number of macroscopically visible plaques; and green bars represent the number of green fluorescent plaques that were counted using a fluorescence microscope. N > 5; error bars show the means ± SEM. ND, not detected. (G) An isolated green HSV1 plaque that was observed using a fluorescence microscope and merged with an image of DAPI staining. Scale bar, 50 µm. (H) PCR amplification of HSV1 genomes extracted from a green fluorescent plaque using oligo 8 and oligo 9 to verify the lack of wild-type HSV1 contamination.

    Article Snippet: Restriction fragment length polymorphism (RFLP) analysis of gene modifications Purified HSV1 DNA products from the genomic DNA extraction and PCR amplification were digested with BsiWI (New England Biolabs) for 16 hours at 55°C and analyzed on 0.5 µg/ml ethidium bromide-stained agarose gels (1%).

    Techniques: CRISPR, Amplification, RFLP Assay, Expressing, Infection, Generated, Titration, Endpoint Dilution Assay, Recombinant, Plaque Assay, Fluorescence, Microscopy, Isolation, Staining, Polymerase Chain Reaction