人转录因子Slug过表达慢病毒载体的构建与鉴定

摘要

Objective To construct recombinant lentivirus with Slug gene for further exploring the effect of Slug in metastatic recurrence.Methods The Slug gene was acquired from the breast cancer cell MDA-MB-231 by PCR assay,which was then connected into linearized GV367 vector digested by Age Ⅰ/Nhe Ⅰ enzyme.The positive clones were identified by PCR and sequencing analysis.The constructed lentiviral vectors with Slug gene were then transfered into 293T cell lines.The lentivirus titer was detected by the fluorescence method.Results The lentiviral vector with Slug gene was successfully constructed and obtained,the virus titer of which was 5×108 TU/mL.Conclusion The lentiviral with Slug gene are successfully constructed,which will provide the experimental foundation for further exploring the effect of Slug in metastatic recurrence.%目的 构建Slug过表达的慢病毒载体,为探讨Slug在肿瘤复发转移中的影响提供前期基础.方法 从人乳腺癌细胞MDA-MB-231中获得Slug基因片段,与Age Ⅰ/Nhe Ⅰ酶切后成线性化的GV367载体连接,采用PCR及DNA测序的方式鉴定阳性克隆.将构建的LV-Slug转染入293T细胞中,采用荧光法检测LV-Slug的病毒滴度.结果 经测序证实Slug基因已成功插入到病毒载体中,其病毒滴度为5×108 TU/mL.结论成功构建了携带Slug过表达的慢病毒载体,为探讨Slug在肿瘤转移复发中的影响提供基础.