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  • Title: [Genetic system of selecting point mutations of bacteriophage T7 RNA polymerase].
    Author: Rechinskiĭ VO, Dragan SM, Kostiuk DA, Liakhov DL, Kochetkov SN.
    Journal: Mol Biol (Mosk); 1991; 25(6):1588-93. PubMed ID: 1813803.
    Abstract:
    Random mutagenesis of the bacteriophage T7 gene 1 was used to delineate the functionally important amino acids residues of its product--T7 RNA polymerase. A two-plasmid system has been constructed for the phenotypic selection of the mutants. Nine mutants were isolated; the RNA polymerase with Tyr-639----Asp substitution has been shown to be completely inactive. Methods were developed for the rapid purification of plasmid-encoded mutant proteins, facilitating their future biochemical analysis.
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