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. 1974 Mar;71(3):760–764. doi: 10.1073/pnas.71.3.760

A Preliminary Map of the Major Transcription Units Read by T7 RNA Polymerase on the T7 and T3 Bacteriophage Chromosomes*,†

Miriam Golomb 1,2, Michael Chamberlin 1,2
PMCID: PMC388093  PMID: 4522790

Abstract

Transcription of T7 DNA by T7 RNA polymerase in vitro gives rise to six major size classes of RNAs comprising seven major T7 RNA species. These RNAs are all read from the r-strand of T7 DNA and are not derived from the early (leftmost on the conventional genetic map) region of the molecule. When artifically shortened T7 DNA templates are transcribed, four (I, II, IIIb, and VI) of the seven species are found to be truncated or deleted. This indicates that all are terminated near the right end of the T7 DNA molecule, probably at a common termination site near 98.5%. (Map positions are all given in terms of percentage of total length measured from the left end of the molecule.) Since the approximate lengths of the transcripts are known, the promotor sites for T7 RNA species I, II, IIIb, and VI are tentatively mapped at 56, 64, 83, and 97% on the T7 chromosome. Only a single major T3 RNA is transcribed by T7 RNA polymerase; analysis of transcripts directed by shortened T3 DNA templates indicates it is analogous to T7 RNA species IIIb. Hence the promotor and terminator sites for T3 species IIIb are tentatively mapped at 83 and 98.5%, respectively, on the T3 chromosome. The major transcripts read by T7 RNA polymerase from T3-T7 hybrid phage DNAs vary, depending on which regions of the T7 chromosome are present. This provides an alternative method of mapping the strong T7 promotor sites on the T7 chromosome.

Keywords: in vitro T7 RNAs, gel electrophoresis, exonuclease cleavage, T3-T7 hybrid phages

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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