c-myc基因慢病毒载体的构建及其意义

目的 构建携带c-myc基因野生型及T58A突变型的慢病毒载体.方法 应用聚合酶链反应(PCR)方法 扩增c-myc野生型及T58A突变型基因,利用Gateway技术构建携带绿色荧光蛋白(GFP)基因的慢病毒载体,经PCR及基因测序鉴定后,转染293FT包装细胞,产生相应慢病毒,测定其滴度.结果 PCR和测序证实,构建分别携带c-myc野生型及T58A突变型基因的慢病毒载体,并包装慢病毒,病毒滴度测定结果　分别为6.10 × 107、5.65×107 TU/ml.结论　成功构建含有c-myc野生型及T58A突变型基因的慢病毒载体并包装出具高效感染力的慢病毒颗粒.

Abstract：

Objective To construct a lentiviral vector with c-myc gene, including wild type and mutation type. Methods The lentiviral vector with green fluorescent protein (GFP) was constructed by polymerase chain reaction (PCR) and gateway technology and identified by PCR and gene sequencing,then transfecte dinto the package cells 293FT by lipofectin to produce mature lentivirus. The virus titer was measured. Results According to PCR and gene sequencing, the myc-lentiviral vectors were successfully constructed with the virus titer being 6. 10 × 107 ( wild type) and 5.65 × 107 ( mutation type) TU/ml, respectively. Conclusion The myc-lentiviral vector was successfully constructed and efficient lentivirus particles were packaged.

Construction and significance of lentiviral vector with c-myc

Objective To construct a lentiviral vector with c-myc gene, including wild type and mutation type. Methods The lentiviral vector with green fluorescent protein (GFP) was constructed by polymerase chain reaction (PCR) and gateway technology and identified by PCR and gene sequencing,then transfecte dinto the package cells 293FT by lipofectin to produce mature lentivirus. The virus titer was measured. Results According to PCR and gene sequencing, the myc-lentiviral vectors were successfully constructed with the virus titer being 6. 10 × 107 ( wild type) and 5.65 × 107 ( mutation type) TU/ml, respectively. Conclusion The myc-lentiviral vector was successfully constructed and efficient lentivirus particles were packaged.