Inglis M M, Darby G
J Gen Virol. 1987 Jan;68 ( Pt 1):39-46. doi: 10.1099/0022-1317-68-1-39.
The thymidine kinase (TK) gene from herpes simplex virus type 1 strain SC16 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.
将单纯疱疹病毒1型SC16株的胸苷激酶（TK）基因克隆到噬菌体M13 mp8中，以便在感染重组噬菌体M13/TK的细菌中表达功能性HSV-1 TK。然后采用寡核苷酸定点诱变技术，将M13/TK中TK基因的单个核苷酸进行改变，从而将野生型酶中半胱氨酸171的密码子替换为编码丝氨酸或甘氨酸的密码子。对细菌提取物中的突变酶进行分析表明，这些替换对酶的活性影响很小，这表明该残基的侧链不参与核苷结合，对酶的催化活性也不是必需的。
