单纯疱疹病毒2型DNA提取及ICP10启动子基因获得和纯化

Isolation of DNA from HSV-2 and Purification of ICP10 Promoter Gene

目的用单纯疱疹病毒2型DNA,扩增并纯化得到ICP10启动子基因。方法培养vero细胞并接种HSV-2,获得大量HSV-2。用酚:氯仿提取病毒DNA,用PCR方法获得病毒ICP10启动子基因,并用UNIQ-10柱试剂盒纯化PCR产物。结果接种病毒的vero细胞出现病变,病变为“”,收集细胞,反复冻融3次,使细胞裂解,获得大量HSV-2。经PCR扩增获得641bp大小的目的基因片段。电泳显示片断大小符合。结论获得的ICP10启动子基因,经过测序正确后,可以和其他基因连接,用于进一步研究。

Objective Amplification, purification of ICP10 promoter gene. Methods DNA of HSV- 2 is isolated from culture vero cells.A pair of PCR primers were created from the ICP10 promoter gene of HSV-2 （Sal 5 strain） by DNA Star software and two restriction sites （Bgl Ⅱ and Hind Ⅲ ） were included as part of the primers, The gene fragment coding for ICP10 promoter was amplified by PCR. The PCR product was purified by PCR purification kit. Results A 641bp fragment was amplified by PCR. The gene fragment was proved to correct by agarose gel electrophoresis. Conclusion The gene fragment is used to ligate to vector after DNA sequencing.