T7 RNA Polymerase promoter question

Keith Robison robison at mito.harvard.edu
Wed Aug 24 16:41:40 EST 1994


jr at dna.bio.warwick.ac.uk wrote:
: Dear all,

:       Maybe someone out there can help me with this question regarding the
: T7 RNA promoter. So far as I can find out the promoter sequence is:
: (From the Stratagene catalog)


: TAATACGACTCACTATA *GGG

:  in which transcription initiates at the first G residue of the three G's and 
: is marked with an asterix

: My question is : Do those three G's have to be there or could I replace the
: three G's without loss of transcription activity?

: I would ideally like to replace them with TGC. This is so as to intiate 
: transcription at a start of my insert. (Before someone posts asking why I'm 
: doing this since translation would occur further down and therefore I 
: don't need to change the T7 promoter sequence, let me say its just a long 
: story and the work necessitates this change. If you really want to know I 
: don't mind E-mailing responses.)


Tom Schneider did a randomization study on T7 RNAP sites which 
is quite interesting; you will probably find a suitable answer there.

	Excess information at bacteriophage T7 genomic promoters detected by
	a random cloning technique.  Nuc.Acids.Res 17:659-674

	See also:
		Schneider et al.  JMB 188:415-431

Enjoy & good luck!


Keith Robison
Harvard University
Department of Cellular and Developmental Biology
Department of Genetics / HHMI

robison at mito.harvard.edu 





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