Mutant T7 RNA polymerase GP1(lys222) exhibiting altered promoter

Details Mutant T7 RNA polymerase GP1(lys222) exhibiting altered promoter Mutant T7 RNA polymerase GP1(lys222) exhibiting altered promoter

1993-08-13

1995-01-31

Baker, Keith

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

435194, 536 232, 935 31, 935 41, C07H 2100, C12N 912, C12N 1554

Patent

active

053858344

ABSTRACT:
E. coli harboring the mutant plasmid pKGP-HA1mut4 and an inactive pCM-X# are chloramphenicol resistant and that the mutation responsible for the expression of CAT from the inactive pCM-X# plasmid is a G to A transition at nucleotide 664 of T7 gene 1 that converts glutamic acid (222) to lysine. This mutation expands the range of T7 promoter sequences that can be utilized by the enzyme. The mutant T7 RNA polymerase, GP1(lys222), utilizes inactive T7 promoter point mutants more efficiently than wild-type T7 RNA polymerase both in vivo and in vitro. Furthermore, the correlation of in vivo and in vitro promoter utilization suggests that the restoration of chloramphenicol resistance in the cotransformed E. coli results from the ability of GP1(lys222) to initiate transcription from T7 promoter point mutants that are normally inactive.

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