Mutants of T7 RNA polymerase that are able to synthesize both RNA and DNA

FEBS Letters

Chernov, B.K.¹  Dragan, S.M.¹  Tunitskaya, V.L.¹  Kostyuk, D.A.¹  Lyakhov, D.L.¹  Rechinsky, V.O.¹  Kochetkov, S.N.¹ 

[1] Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov st. 32, 117984 Moscow, Russian Federation

关键词: T7 RNA polymerase;    Mutagenesis;    dNTP utilization;    DNA polymerizing activity;   

DOI  :  10.1016/0014-5793(95)00732-O

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown