NDLI: Relationship between promoter structure and template specificities exhibited by the bacteriophage T3 and T7 RNA polymerases.

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Relationship between promoter structure and template specificities exhibited by the bacteriophage T3 and T7 RNA polymerases.

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Relationship between promoter structure and template specificities exhibited by the bacteriophage T3 and T7 RNA polymerases.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Klement, J. F. Bailey, J. N. Mcallister, W. T.
Abstract	To explore the basis for the template specificities of the bacteriophage T3 and T7 RNA polymerases (EC 2.7.7.6), we determined the nucleotide sequences of six promoters recognized by the T3 RNA polymerase and compared them with the previously determined promoter sequences recognized by the bacteriophage T7 RNA polymerase. Recombinant plasmids containing random Hpa II and Taq I fragments of T3 DNA were screened for T3 promoter activity in vitro in a transcription assay using purified T3 RNA polymerase. Five promoters for the T3 RNA polymerase were identified in this manner and their sequences were determined; the sequence of an additional promoter was determined directly from a genomic DNA fragment. In five of the T3 promoters an identical 16-base-pair sequence (A-C-C-C-T-C-A-C-T-A-A-A-G-G-G-A) extends from -12 to +4 (initiation occurring with GTP at +1); this sequence is preceded by a 6-base-pair A + T region. The remaining promoter contains an inserted C at position -1 and an A at the +1 position. The sequence of the 5' end of the RNA transcript from the latter promoter confirms that transcription is initiated with ATP at the +1 position. Previously, late T3 or T7 transcripts had not been found to initiate with ATP. The highly conserved T3 promoter sequence was compared to the T7 promoter consensus sequence. The fundamental difference between the two kinds of phage promoters is the occurrence of G-A at positions -11 and -10 in the T7 promoter, whereas there is a single C at position -10 in the T3 promoter.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	10
Volume Number	80
Language	English
Publisher	National Academy of Sciences
Publisher Date	1983-07-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA-Directed RNA Polymerases Metabolism Operon T-Phages Enzymology RNA, Viral Biosynthesis Substrate Specificity Transcription, Genetic Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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