Analysis of the Role of the Cysteine 171 Residue in the Activity of Herpes Simplex Virus Type 1 Thymidine Kinase by Oligonucleotide-directed Mutagenesis
The thymidine kinase (TK) gene from herpes simplex virus type 1 strain SC 16 was cloned into bacteriophage M13 mp8 so that functional HSV-1 TK was expressed in bacteria infected with the recombinant bacteriophage, M13/TK. Oligonucleotide site- directed mutagenesis was then employed to introduce single nucleotide changes into the TK gene in M13/TK in order to alter the codon for cysteine 171 in the wild-type enzyme to a codon specifying either serine or glycine. Analysis of the mutant enzymes in bacterial extracts showed that these substitutions had little effect on the activity of the enzyme, indicating that the side chain of this residue is not involved in nucleoside binding and is not essential for the catalytic activity of the enzyme.
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Analysis of the Role of the Cysteine 171 Residue in the Activity of Herpes Simplex Virus Type 1 Thymidine Kinase by Oligonucleotide-directed Mutagenesis