猕猴肉瘤病毒SV40 T-ag-C 基因克隆及序列分析

摘要: 目的对来源于一株自然感染猕猴肉瘤病毒SV40 的猴肾细胞培养物进行大T 抗原C-羧基端( T-ag-C) 基因 克隆及核苷酸序列分析。方法采用PCR 法分别从一株自然感染SV40 病毒的猴肾细胞培养物和SV40776 标准 株接种的vero 细胞培养物提取的总DNA 中扩增出441bp 的SV40 大T 抗原C-羧基端( T-ag-C) 基因片段，分别将其 克隆到PMD18-T 载体中，转化至JM109 感受态大肠杆菌细胞后，挑取阳性克隆进行测序鉴定，并对获得的目的基 因核苷酸序列进行序列分析及同源性分析。结果来源于猴肾细胞培养物的SV40 大T 抗原片段与本实验室来源 于云南野生猴群的猕猴外周血所得到的SV40 大T 抗原片段同源性为97. 31% ，与GenBank 中登录号为NC _ 001669. 1 序列进行比对，同源性为96. 33% ; 与SV40-776 标准株接种的vero 细胞培养物所扩增的大T 抗原片段同 源性为97. 55% 。结论对大T 抗原基因克隆和序列分析是了解和掌握SV40 病毒分子流行病学及其变异趋势的 重要手段。

Clone and Sequence Analysis of the T-ag-C Gene of Simian Virus 40 from a Kidney Culture of Macaca monkey

Abstract: Objective To clone of the large tumor antigen( T-ag-C) gene of simian virus 40 from kidney culture of Macaca monkey and analyze its sequence． Method The 441 bp DNA segment were obtained by PCR from both kidney culture of Macaca monkey and culture of SV40 strain 776 using primers T-ag-C gene of simian virus 40，and they were cloned into the vector PMD18-T for sequencing． The T-ag-C gene from kidney culture of Macaca monkey and culture of SV40 strain 776 were identified by comparison with other simian viruses 40 from gene bank and other reference． Result The sequence comparison with the T-ag-C gene of SV40 Macaca-RMK． seq showed nucleotide homologies of 97. 31% with SV40-Macaca-PBMC． seq，96. 33% with gene bank SV40-GI-9628421. seq， 97. 55% with SV40-strain-776. seq． Conclusion Sequencing DNA of gene is a powerful tool for classification as well as molecular epidemiology of simian virus 40 ．