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目的：对单纯疱疹Ⅰ型胸苷激酶基因扩增、克隆和测序．方法：从单纯疱疹病毒Ⅰ型（ＨＳＶ－１）１７ｓｙｎ＋株感染的ＢＨＫ２１细胞上清液中提取ＨＳＶ－１基因组ＤＮＡ，以此为模板，用ＰＣＲ方法扩增胸苷激酶（ＴＫ）．将扩增的片段克隆入载体ｐＵＣ１８中，并进行测序．结果：除终止码外，ＨＳＶ－１ＴＫ基因全部编码区为１１２８ｂｐ，编码３７６个氨基酸，其中含１２个蛋氨酸，４个半胱氨酸．ＴＫ的第２４９和２５０位氨基酸残基分别为Ｌｅｕ和Ｇｌｎ，其相应密码子为ＣＴＧ，ＧＡＧ，构成１个ＰｓｔＩ位点，第３１３和３１４氨基酸残基分别为Ａｓｐ和Ｖａｌ，其相应密码子为ＧＡＣ和ＧＴＣ，构成另一个ＰｓｔＩ位点．结论：本研究扩增出了ＨＳＶ－１ＴＫ基因的全部编码区序列
Objective: To amplify, clone and sequence the herpes simplex type Ⅰ thymidine kinase gene. Methods: HSV-1 genomic DNA was extracted from BHK21 cell supernatant infected with herpes simplex virus type 1 (HSV-1) 17syn + strain and used as template to amplify thymidine kinase (TK) by PCR. The amplified fragment was cloned into the vector pUC18 and sequenced. Results: Except for the stop codon, the entire coding region of HSV-1TK gene was 1128bp, encoding 376 amino acids, including 12 methionine and 4 cysteines. The amino acid residues at positions 249 and 250 of TK are Leu and Gln, respectively, and the corresponding codons are CTG and GAG, forming one PstI site. The amino acid residues 313 and 314 are respectively Asp and Val, and the corresponding codons are GAC and GTC, constitute another PstI site. Conclusion: This study amplified all the coding sequence of HSV-1TK gene