Difference between revisions of "Part:BBa I20530"
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"The consensus promoter sequences for SP6 and K1-5 differ at positions −10, −11, −12 (Table 2 and Table 3). The SP6 consensus −12G and −10T are not major determinants of promoter activity in vivo or in vitro but any substitution at −11G is inactivating.34 All K1-5 promoters have a pyrimidine at −11, suggesting that SP6 RNAP will not transcribe K1-5 DNA and vice versa. A comparable specificity of the phage RNAP for its cognate promoters is seen throughout the T7 group; for example, T7 RNAP does not recognize T3 promoters and vice versa. However, a single amino acid change in T7 RNAP is sufficient to switch its specificity to T3 promoters, and a change at −11 in a T7 promoter is sufficient to allow its recognition by T3 RNAP.[36.] and [37.] Two changes at positions −9 and −8 of the SP6 promoter to those of a T7 promoter allow recognition by T7 RNAP in vitro.38 All SP6 and most K1-5 promoters have the sequence 5′-GA at these positions, suggesting that the K1-5 genome cannot be transcribed efficiently by T7 RNAP." | "The consensus promoter sequences for SP6 and K1-5 differ at positions −10, −11, −12 (Table 2 and Table 3). The SP6 consensus −12G and −10T are not major determinants of promoter activity in vivo or in vitro but any substitution at −11G is inactivating.34 All K1-5 promoters have a pyrimidine at −11, suggesting that SP6 RNAP will not transcribe K1-5 DNA and vice versa. A comparable specificity of the phage RNAP for its cognate promoters is seen throughout the T7 group; for example, T7 RNAP does not recognize T3 promoters and vice versa. However, a single amino acid change in T7 RNAP is sufficient to switch its specificity to T3 promoters, and a change at −11 in a T7 promoter is sufficient to allow its recognition by T3 RNAP.[36.] and [37.] Two changes at positions −9 and −8 of the SP6 promoter to those of a T7 promoter allow recognition by T7 RNAP in vitro.38 All SP6 and most K1-5 promoters have the sequence 5′-GA at these positions, suggesting that the K1-5 genome cannot be transcribed efficiently by T7 RNAP." | ||
| − | == Property Rights == | + | === Property Rights === |
All rights not already held by others are '''PUBLIC DOMAIN''' in compliance with section 5.1.A.3 of the Stanford Research Policy Handbook. http://rph.stanford.edu/5-1.html | All rights not already held by others are '''PUBLIC DOMAIN''' in compliance with section 5.1.A.3 of the Stanford Research Policy Handbook. http://rph.stanford.edu/5-1.html | ||
Revision as of 17:11, 17 June 2011
Consensus promoter for phage K1-5 RNA polymerase
Useful for orthogonal transcription with cognate K1-5 RNA polymerase.
From Scholl et al. Journal of Molecular Biology, Volume 335, Issue 5, 30 January 2004, Pages 1151-1171
"Most SP6 and K1-5 transcription occurs using their respective phage-encoded RNAPs. Several SP6 promoters have been identified and cloned and transcriptional activity with respect to promoter sequence has been examined comprehensively by saturation mutagenesis.[33.] and [34.] On the basis of this information we have identified 12 phage RNAP promoters in the SP6 genome. Only the promoter upstream of ORF31 conforms exactly to the consensus. Three promoter-like sequences that contain one or more nucleotide changes at conserved positions, including+1G that is required for SP6 RNAP transcription initiation in vitro, were found but are likely not to be functional."
"The consensus promoter sequences for SP6 and K1-5 differ at positions −10, −11, −12 (Table 2 and Table 3). The SP6 consensus −12G and −10T are not major determinants of promoter activity in vivo or in vitro but any substitution at −11G is inactivating.34 All K1-5 promoters have a pyrimidine at −11, suggesting that SP6 RNAP will not transcribe K1-5 DNA and vice versa. A comparable specificity of the phage RNAP for its cognate promoters is seen throughout the T7 group; for example, T7 RNAP does not recognize T3 promoters and vice versa. However, a single amino acid change in T7 RNAP is sufficient to switch its specificity to T3 promoters, and a change at −11 in a T7 promoter is sufficient to allow its recognition by T3 RNAP.[36.] and [37.] Two changes at positions −9 and −8 of the SP6 promoter to those of a T7 promoter allow recognition by T7 RNAP in vitro.38 All SP6 and most K1-5 promoters have the sequence 5′-GA at these positions, suggesting that the K1-5 genome cannot be transcribed efficiently by T7 RNAP."
Property Rights
All rights not already held by others are PUBLIC DOMAIN in compliance with section 5.1.A.3 of the Stanford Research Policy Handbook. http://rph.stanford.edu/5-1.html
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]