Bacteriophage SP6-specific RNA polymerase. II. Mapping of SP6 DNA and selective in vitro transcription.
نویسندگان
چکیده
منابع مشابه
Sequences of three promoters for the bacteriophage SP6 RNA polymerase.
Fragments of SP6 DNA generated by cleavage with Hpa II or Taq I were cloned into the Cla I site of pBR322 and the recombinant plasmids were screened for the presence of SP6 promoter activity by transcription in vitro with purified SP6 RNA polymerase. Three plasmids having promoter activity and small inserts of SP6 DNA were characterized. Hybridization studies showed that all three cloned promot...
متن کاملNucleotide sequence and expression of the cloned gene of bacteriophage SP6 RNA polymerase.
The coding region of the gene for bacteriophage SP6 RNA polymerase was cloned into pBR322, and its entire nucleotide sequence was deduced. The predicted amino acid sequence for the polymerase consists of 874 amino acid residues with a total molecular weight of 98,561 daltons. Comparison of the amino acid sequence with that of T7 RNA polymerase reveals that regions with partial homology are pres...
متن کاملSuitability of SP6 RNA polymerase transcripts for in vitro viral diagnosis.
To the Editor: Application of DNA or RNA probes to microbiology and virology requires probes free of vector sequences, particularly vectors of bacterial origin, such as pBR322. Incomplete purification of inserts from vector sequences can produce a falsely positive result when nucleic acid probes are applied to tissue or body fluids that may be containmated with bacteria. A plasrnid vector such ...
متن کاملTranscription initiation site selection and abortive initiation cycling of phage SP6 RNA polymerase.
Effects of mutations around the phage SP6 transcription initiation site on SP6 RNA polymerase's selection of initiation site were studied. In the in vitro transcription reactions, the limiting concentration of a ribonucleotide causes the SP6 RNA polymerase to stall long enough only at the positions of the limited nucleotide and dissociate from the elongation complex. As a result, a series of RN...
متن کاملSP6 RNA polymerase efficiently synthesizes RNA from short double-stranded DNA templates.
SP6 DNA-dependent RNA polymerase, like T7 RNA polymerase, can be used to synthesize RNA sequences from short DNA templates which contain the 18 base pair promoter region. Use of SP6 polymerase extends the range of possible 5' sequences of RNA products, since the preferred SP6 start site (of the RNA product) is 5'GAAGA, while T7 polymerase prefers 5'GGGAG. The SP6 start site can be advantageous ...
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ژورنال
عنوان ژورنال: Journal of Biological Chemistry
سال: 1982
ISSN: 0021-9258
DOI: 10.1016/s0021-9258(19)83847-4