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Mutants of T7 RNA polymerase that are able to synthesize both RNA and DNA
Author(s) -
Kostyuk D.A.,
Dragan S.M.,
Lyakhov D.L.,
Rechinsky V.O.,
Tunitskaya V.L.,
Chernov B.K.,
Kochetkov S.N.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00732-o
Subject(s) - t7 rna polymerase , terminator (solar) , polymerase , primase , microbiology and biotechnology , rna dependent rna polymerase , rna polymerase , primer (cosmetics) , dna polymerase , dna , dna clamp , biology , dna polymerase ii , rna , mutant , rna polymerase i , biochemistry , chemistry , gene , bacteriophage , reverse transcriptase , escherichia coli , ionosphere , physics , organic chemistry , astronomy
A mutant T7 RNA polymerase (T7 RNAP) having two amino‐acid substitutions (Y639F and S641A) is altered in its specificity towards nucleotide substrates, but is not affected in the specificity of its interaction with promoter and terminator sequences. The mutant enzyme gains the ability to utilize dNTPs and catalyze RNA and DNA synthesis from circular supercoiled plasmid DNA. DNA synthesis can also be initiated from a single stranded template using a DNA primer. Another T7 RNAP mutant having only the single substitution S641A loses RNA polymerase activity but is able to synthesize DNA.