Structured Review

5 PRIME primer sequences
Primer Sequences, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/primer sequences/product/5 PRIME
Average 92 stars, based on 1 article reviews
Price from $9.99 to $1999.99
primer sequences - by Bioz Stars, 2020-07
92/100 stars

Images

Related Articles

Countercurrent Chromatography:

Article Title: Zika virus replicates in adult human brain tissue and impairs synapses and memory in mice
Article Snippet: .. Primer sequences were the following: IL-1β Forward: 5prime;-GTA ATG AAA GAC GGC ACA CC-3′ and Reverse: 5′-ATT AGA AAC AGT CCA GCC CA-3′; TNF-α Forward: 5′-CCC TCA CAC TCA GAT CAT CTT CT-3′ and Reverse: 5′-GCT ACG ACG TGG GCT ACA G-3′; Actin Forward: 5′-TGT GAC GTT GAC ATC CGT AAA-3′ and Reverse: 5′-GTA CTT GCG CTC AGG AGG AG-3′; C1q Forward: 5′-CTC AGG GAT GGC TGG TGG CC-3′ and Reverse: 5′-CCT TTG AGA CCC GGC CTC CCC-3′; C3 Forward: 5′-ACC CAG CTC TGT GGG AAG TG-3′ and Reverse: 5′-CTT CAT AGA CTG CTG CAA CCA-3′. .. Immunohistochemistry Mice were transcardially perfused with cold PBS followed by ice-cold 4% paraformaldehyde.

CTG Assay:

Article Title: Zika virus replicates in adult human brain tissue and impairs synapses and memory in mice
Article Snippet: .. Primer sequences were the following: IL-1β Forward: 5prime;-GTA ATG AAA GAC GGC ACA CC-3′ and Reverse: 5′-ATT AGA AAC AGT CCA GCC CA-3′; TNF-α Forward: 5′-CCC TCA CAC TCA GAT CAT CTT CT-3′ and Reverse: 5′-GCT ACG ACG TGG GCT ACA G-3′; Actin Forward: 5′-TGT GAC GTT GAC ATC CGT AAA-3′ and Reverse: 5′-GTA CTT GCG CTC AGG AGG AG-3′; C1q Forward: 5′-CTC AGG GAT GGC TGG TGG CC-3′ and Reverse: 5′-CCT TTG AGA CCC GGC CTC CCC-3′; C3 Forward: 5′-ACC CAG CTC TGT GGG AAG TG-3′ and Reverse: 5′-CTT CAT AGA CTG CTG CAA CCA-3′. .. Immunohistochemistry Mice were transcardially perfused with cold PBS followed by ice-cold 4% paraformaldehyde.

Chloramphenicol Acetyltransferase Assay:

Article Title: Zika virus replicates in adult human brain tissue and impairs synapses and memory in mice
Article Snippet: .. Primer sequences were the following: IL-1β Forward: 5prime;-GTA ATG AAA GAC GGC ACA CC-3′ and Reverse: 5′-ATT AGA AAC AGT CCA GCC CA-3′; TNF-α Forward: 5′-CCC TCA CAC TCA GAT CAT CTT CT-3′ and Reverse: 5′-GCT ACG ACG TGG GCT ACA G-3′; Actin Forward: 5′-TGT GAC GTT GAC ATC CGT AAA-3′ and Reverse: 5′-GTA CTT GCG CTC AGG AGG AG-3′; C1q Forward: 5′-CTC AGG GAT GGC TGG TGG CC-3′ and Reverse: 5′-CCT TTG AGA CCC GGC CTC CCC-3′; C3 Forward: 5′-ACC CAG CTC TGT GGG AAG TG-3′ and Reverse: 5′-CTT CAT AGA CTG CTG CAA CCA-3′. .. Immunohistochemistry Mice were transcardially perfused with cold PBS followed by ice-cold 4% paraformaldehyde.

Cellular Antioxidant Activity Assay:

Article Title: Zika virus replicates in adult human brain tissue and impairs synapses and memory in mice
Article Snippet: .. Primer sequences were the following: IL-1β Forward: 5prime;-GTA ATG AAA GAC GGC ACA CC-3′ and Reverse: 5′-ATT AGA AAC AGT CCA GCC CA-3′; TNF-α Forward: 5′-CCC TCA CAC TCA GAT CAT CTT CT-3′ and Reverse: 5′-GCT ACG ACG TGG GCT ACA G-3′; Actin Forward: 5′-TGT GAC GTT GAC ATC CGT AAA-3′ and Reverse: 5′-GTA CTT GCG CTC AGG AGG AG-3′; C1q Forward: 5′-CTC AGG GAT GGC TGG TGG CC-3′ and Reverse: 5′-CCT TTG AGA CCC GGC CTC CCC-3′; C3 Forward: 5′-ACC CAG CTC TGT GGG AAG TG-3′ and Reverse: 5′-CTT CAT AGA CTG CTG CAA CCA-3′. .. Immunohistochemistry Mice were transcardially perfused with cold PBS followed by ice-cold 4% paraformaldehyde.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • utr  (5 PRIME)
    92
    5 PRIME utr
    PCA of gene features grouped by <t>MSL</t> binding and expression status . Results of PCA of gene feature sequences of genes that are expressed/unexpressed and strongly/weakly bound by MSL. Chromosomes colour-coded, and gene features indicated by symbols as follows: green = X, magenta = 3R, brown = 2R, blue = 3L, yellow = 2L; O = all genes, Δ = expressed genes (EXP), ∇ = unexpressed genes (UN-EXP), + = MSL strongly bound genes (MSL-SB), × = MSL weakly bound genes (MSL-WB), Δ = expressed MSL strongly bound genes (EXP-MSL-SB), ∇ = expressed MSL weakly bound genes (EXP-MSL-WB). (A) , (B) , (C) , (D) , (E) and (F) show 1 st vs 2 nd component score plots (R2cum = 0.657, 0.517, 0.732, 0.872, 0.577 and 0.761, respectively) of the AT-normalized 2-6 mer promoter, 5' <t>UTR,</t> coding sequence, intron, 3' UTR and intergenic sequence analyses, respectively.
    Utr, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/utr/product/5 PRIME
    Average 92 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    utr - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    88
    5 PRIME 5 prime portion
    Schematic diagram of strand-specific library synthesis mechanism . mRNAs are fragmented by heat and magnesium (1) and primed for cDNA synthesis by an adapter-containing oligonucleotide (2,3) . Size selection and cleanup removes unincorperated oligonucleotides and small cDNA fragments (4) . Transient duplex breathing at the terminus of the RNA-cDNA hybrid (5) facilitates interaction with the single-stranded portion of the <t>5-prime</t> capturing adapter (6) and E. coli DNA Polymerase I catalyses its incorporation into a complete library molecule (7) .
    5 Prime Portion, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5 prime portion/product/5 PRIME
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    5 prime portion - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    88
    5 PRIME dicer1 rnase iiib domain mutations
    Somatic chromosome 14 copy-number segments and allele fractions of variants heterozygous in matched normal. Data points are the tumor allele fractions of heterozygous, non-reference alleles in the matched normal sample. Somatic deletion of the non-reference allele results in tumor allele fractions near 0, whereas gain of the non-reference base or deletion of the reference base results in values approaching 1. Colored bars represent copy-number states inferred from fractional coverage values. Each panel depicts different combinations of copy-number alteration and loss-of-heterozygosity detected by whole exome sequencing. ( a ) Copy quiet : Compound germline loss-of-function (LOF) and somatic <t>RNase</t> <t>IIIb</t> missense mutation without further copy-number alteration (nine cases). ( b ) Wild-type deletion : Deletion of wild-type allele resulting in hemizygosity for the somatic RNase IIIb missense mutation (PPB_11). ( c ) Trisomy : Copy-number gain of chromosome 14 resulting in duplication of RNase IIIb mutant allele and retention of germline LOF allele (two cases). An additional case has duplication of the germline LOF allele and retention of the RNase IIIb mutant allele (PPB_5). ( d ) Chromosomal copy neutral loss-of-heterozygosity : Copy-neutral loss of wild-type allele and duplication of entire chromosome 14 containing the somatic RNase IIIb missense mutation (PPB_15). ( e ) Arm-level copy neutral loss-of-heterozygosity : Copy-neutral loss of wild-type allele and duplication of 14q containing the somatic RNase IIIb missense mutation (PPB_13).
    Dicer1 Rnase Iiib Domain Mutations, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dicer1 rnase iiib domain mutations/product/5 PRIME
    Average 88 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    dicer1 rnase iiib domain mutations - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    88
    5 PRIME gpt gene
    Construction of a vaccinia virus based <t>SARS-CoV</t> reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and <t>gpt</t> PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.
    Gpt Gene, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gpt gene/product/5 PRIME
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    gpt gene - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    Image Search Results


    PCA of gene features grouped by MSL binding and expression status . Results of PCA of gene feature sequences of genes that are expressed/unexpressed and strongly/weakly bound by MSL. Chromosomes colour-coded, and gene features indicated by symbols as follows: green = X, magenta = 3R, brown = 2R, blue = 3L, yellow = 2L; O = all genes, Δ = expressed genes (EXP), ∇ = unexpressed genes (UN-EXP), + = MSL strongly bound genes (MSL-SB), × = MSL weakly bound genes (MSL-WB), Δ = expressed MSL strongly bound genes (EXP-MSL-SB), ∇ = expressed MSL weakly bound genes (EXP-MSL-WB). (A) , (B) , (C) , (D) , (E) and (F) show 1 st vs 2 nd component score plots (R2cum = 0.657, 0.517, 0.732, 0.872, 0.577 and 0.761, respectively) of the AT-normalized 2-6 mer promoter, 5' UTR, coding sequence, intron, 3' UTR and intergenic sequence analyses, respectively.

    Journal: BMC Genomics

    Article Title: Sequence signatures involved in targeting the male-specific lethal complex to X-chromosomal genes in Drosophila melanogaster

    doi: 10.1186/1471-2164-13-97

    Figure Lengend Snippet: PCA of gene features grouped by MSL binding and expression status . Results of PCA of gene feature sequences of genes that are expressed/unexpressed and strongly/weakly bound by MSL. Chromosomes colour-coded, and gene features indicated by symbols as follows: green = X, magenta = 3R, brown = 2R, blue = 3L, yellow = 2L; O = all genes, Δ = expressed genes (EXP), ∇ = unexpressed genes (UN-EXP), + = MSL strongly bound genes (MSL-SB), × = MSL weakly bound genes (MSL-WB), Δ = expressed MSL strongly bound genes (EXP-MSL-SB), ∇ = expressed MSL weakly bound genes (EXP-MSL-WB). (A) , (B) , (C) , (D) , (E) and (F) show 1 st vs 2 nd component score plots (R2cum = 0.657, 0.517, 0.732, 0.872, 0.577 and 0.761, respectively) of the AT-normalized 2-6 mer promoter, 5' UTR, coding sequence, intron, 3' UTR and intergenic sequence analyses, respectively.

    Article Snippet: Abbreviations Dm : Drosophila melanogaster ; MSL: Male-specific lethal; CDS: Coding Sequence; 5' UTR: 5 prime untranslated region; 3' UTR: 3 prime untranslated region; TSS: Transcription start site; HAS: High affinity site; PCA: Principal component analysis; OPLS-DA: Orthogonal partial least squares discriminant analysis; PWM: Position weight matrix

    Techniques: Binding Assay, Expressing, Western Blot, Sequencing

    PCA of D. melanogaster gene features . Results of PCA of frequencies of sequence words within gene features in Dm chromosome arms. Chromosomes colour-coded, and gene features indicated by symbols as follows: green = X, magenta = 3R, brown = 2R, blue = 3L, yellow = 2L; ∇ = promoter, Δ = 5'UTR, □ = CDS, O = intron, + = 3'UTR, × = intergenic. (A) Scatter plot of PCA first component scores of the gene features versus their AT contents. (B), (C) and (D): 1 st vs 2 nd , 3 rd vs 4 th and 5 th vs 6 th component score plots (R2cum = 0.774, 0.923 and 0.954, respectively) of the AT-normalized analysis of 2-6 mer frequencies in gene features.

    Journal: BMC Genomics

    Article Title: Sequence signatures involved in targeting the male-specific lethal complex to X-chromosomal genes in Drosophila melanogaster

    doi: 10.1186/1471-2164-13-97

    Figure Lengend Snippet: PCA of D. melanogaster gene features . Results of PCA of frequencies of sequence words within gene features in Dm chromosome arms. Chromosomes colour-coded, and gene features indicated by symbols as follows: green = X, magenta = 3R, brown = 2R, blue = 3L, yellow = 2L; ∇ = promoter, Δ = 5'UTR, □ = CDS, O = intron, + = 3'UTR, × = intergenic. (A) Scatter plot of PCA first component scores of the gene features versus their AT contents. (B), (C) and (D): 1 st vs 2 nd , 3 rd vs 4 th and 5 th vs 6 th component score plots (R2cum = 0.774, 0.923 and 0.954, respectively) of the AT-normalized analysis of 2-6 mer frequencies in gene features.

    Article Snippet: Abbreviations Dm : Drosophila melanogaster ; MSL: Male-specific lethal; CDS: Coding Sequence; 5' UTR: 5 prime untranslated region; 3' UTR: 3 prime untranslated region; TSS: Transcription start site; HAS: High affinity site; PCA: Principal component analysis; OPLS-DA: Orthogonal partial least squares discriminant analysis; PWM: Position weight matrix

    Techniques: Sequencing

    Schematic diagram of strand-specific library synthesis mechanism . mRNAs are fragmented by heat and magnesium (1) and primed for cDNA synthesis by an adapter-containing oligonucleotide (2,3) . Size selection and cleanup removes unincorperated oligonucleotides and small cDNA fragments (4) . Transient duplex breathing at the terminus of the RNA-cDNA hybrid (5) facilitates interaction with the single-stranded portion of the 5-prime capturing adapter (6) and E. coli DNA Polymerase I catalyses its incorporation into a complete library molecule (7) .

    Journal: Frontiers in Plant Science

    Article Title: BrAD-seq: Breath Adapter Directional sequencing: a streamlined, ultra-simple and fast library preparation protocol for strand specific mRNA library construction

    doi: 10.3389/fpls.2015.00366

    Figure Lengend Snippet: Schematic diagram of strand-specific library synthesis mechanism . mRNAs are fragmented by heat and magnesium (1) and primed for cDNA synthesis by an adapter-containing oligonucleotide (2,3) . Size selection and cleanup removes unincorperated oligonucleotides and small cDNA fragments (4) . Transient duplex breathing at the terminus of the RNA-cDNA hybrid (5) facilitates interaction with the single-stranded portion of the 5-prime capturing adapter (6) and E. coli DNA Polymerase I catalyses its incorporation into a complete library molecule (7) .

    Article Snippet: These include, the ligation of a known sequence to the 5-prime portion of mRNA molecules prior to cDNA synthesis (Lister et al., ), removal of the template RNA strand followed by randomly primed 2nd strand synthesis (Armour et al., ), labeling of first or second strand cDNA molecules with dUTP for enzymatic degradation prior to enrichment (Parkhomchuk et al., ) and the use of terminal transferases to add defined nucleotides to the cDNA molecules (Zhu et al., ; Tang et al., ), with each method having advantages and shortcomings (Levin et al., ).

    Techniques: Selection

    Somatic chromosome 14 copy-number segments and allele fractions of variants heterozygous in matched normal. Data points are the tumor allele fractions of heterozygous, non-reference alleles in the matched normal sample. Somatic deletion of the non-reference allele results in tumor allele fractions near 0, whereas gain of the non-reference base or deletion of the reference base results in values approaching 1. Colored bars represent copy-number states inferred from fractional coverage values. Each panel depicts different combinations of copy-number alteration and loss-of-heterozygosity detected by whole exome sequencing. ( a ) Copy quiet : Compound germline loss-of-function (LOF) and somatic RNase IIIb missense mutation without further copy-number alteration (nine cases). ( b ) Wild-type deletion : Deletion of wild-type allele resulting in hemizygosity for the somatic RNase IIIb missense mutation (PPB_11). ( c ) Trisomy : Copy-number gain of chromosome 14 resulting in duplication of RNase IIIb mutant allele and retention of germline LOF allele (two cases). An additional case has duplication of the germline LOF allele and retention of the RNase IIIb mutant allele (PPB_5). ( d ) Chromosomal copy neutral loss-of-heterozygosity : Copy-neutral loss of wild-type allele and duplication of entire chromosome 14 containing the somatic RNase IIIb missense mutation (PPB_15). ( e ) Arm-level copy neutral loss-of-heterozygosity : Copy-neutral loss of wild-type allele and duplication of 14q containing the somatic RNase IIIb missense mutation (PPB_13).

    Journal: Oncogene

    Article Title: Exome sequencing of pleuropulmonary blastoma reveals frequent biallelic loss of TP53 and two hits in DICER1 resulting in retention of 5p-derived miRNA hairpin loop sequences

    doi: 10.1038/onc.2014.150

    Figure Lengend Snippet: Somatic chromosome 14 copy-number segments and allele fractions of variants heterozygous in matched normal. Data points are the tumor allele fractions of heterozygous, non-reference alleles in the matched normal sample. Somatic deletion of the non-reference allele results in tumor allele fractions near 0, whereas gain of the non-reference base or deletion of the reference base results in values approaching 1. Colored bars represent copy-number states inferred from fractional coverage values. Each panel depicts different combinations of copy-number alteration and loss-of-heterozygosity detected by whole exome sequencing. ( a ) Copy quiet : Compound germline loss-of-function (LOF) and somatic RNase IIIb missense mutation without further copy-number alteration (nine cases). ( b ) Wild-type deletion : Deletion of wild-type allele resulting in hemizygosity for the somatic RNase IIIb missense mutation (PPB_11). ( c ) Trisomy : Copy-number gain of chromosome 14 resulting in duplication of RNase IIIb mutant allele and retention of germline LOF allele (two cases). An additional case has duplication of the germline LOF allele and retention of the RNase IIIb mutant allele (PPB_5). ( d ) Chromosomal copy neutral loss-of-heterozygosity : Copy-neutral loss of wild-type allele and duplication of entire chromosome 14 containing the somatic RNase IIIb missense mutation (PPB_15). ( e ) Arm-level copy neutral loss-of-heterozygosity : Copy-neutral loss of wild-type allele and duplication of 14q containing the somatic RNase IIIb missense mutation (PPB_13).

    Article Snippet: DICER1 RNase IIIb domain mutations lead to defective cleavage of 5p miRNAs from the pre-miRNA loop sequence Double-stranded precursor miRNAs (pre-miRNAs) are normally processed by DICER1 into three maturation products: two potentially functional units derived from either the 5-prime (5p) or 3-prime (3p) arm of the precursor (mature 5p and 3p miRNAs) and the hairpin loop.

    Techniques: Sequencing, Mutagenesis

    Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.

    Journal: PLoS ONE

    Article Title: Reverse Genetics of SARS-Related Coronavirus Using Vaccinia Virus-Based Recombination

    doi: 10.1371/journal.pone.0032857

    Figure Lengend Snippet: Construction of a vaccinia virus based SARS-CoV reverse genetic system. The genome structure of SARS-CoV is shown at the top of the figure. Nine cDNA clones produced from the genomic RNA of SARS-CoV isolate HKU-39849 are shown below. The region of the SARS-CoV genome encompassed by each clone is indicated by the nucleotide number (using the recSARS-CoV sequence; GenBank: JN854286) at the beginning and end of each clone. Restriction enzyme sites used to join the clones are shown, with restriction enzymes sites added to the clones shown in bold. The cDNA fragments isolated from the clones and gpt PCR products covering regions of the genome unstable as cDNA clones were ligated with each other and vaccinia virus DNA to produce two vaccinia virus recombinant clones spanning nts 1–20288 and 20272–29727 of the SARS-CoV genome respectively. The first 2012 nts of the former vaccinia virus recombinant was derived from the SARS-CoV isolate Frankfurt-1 (shaded in dark grey). Vaccinia virus mediated homologous recombination was then used to reconstitute the SARS-CoV subgenomic fragments, introducing regions of cDNA that were unstable in E. coli and repairing errors (*) introduced during the cloning process. This resulted in the vaccinia virus clones vSARS-CoV-5prime and vSARS-CoV-3prime. The SARS-CoV cDNA fragments were isolated from the two vaccinia virus recombinants by restriction enzyme digestion and then joined using unique SfiI and BglI sites that had been introduced into the cDNA. The ligated cDNA fragments were used as a template for in vitro transcription using a T7 polymerase promoter introduced at the 5′ end of the SARS-CoV 5′ cDNA clone to produce a RNA transcript representing the SARS-CoV genome.

    Article Snippet: Subsequently, the gpt gene was replaced by the appropriate SARS-CoV cDNA sequences using homologous recombination involving vaccinia virus vSARS-CoV-5prime-gpt and a plasmid DNA that contained a shorter region of SARS-CoV cDNA (nts 10808 to 12837).

    Techniques: Clone Assay, Produced, Sequencing, Isolation, Polymerase Chain Reaction, Recombinant, Derivative Assay, Homologous Recombination, In Vitro