中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
4期
543-546,后插2
,共5页
c-myc基因%慢病毒%Gateway技术
c-myc基因%慢病毒%Gateway技術
c-myc기인%만병독%Gateway기술
C-myc gene%Lentivirus%Gateway technology
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.
目的 構建攜帶c-myc基因野生型及T58A突變型的慢病毒載體.方法 應用聚閤酶鏈反應(PCR)方法 擴增c-myc野生型及T58A突變型基因,利用Gateway技術構建攜帶綠色熒光蛋白(GFP)基因的慢病毒載體,經PCR及基因測序鑒定後,轉染293FT包裝細胞,產生相應慢病毒,測定其滴度.結果 PCR和測序證實,構建分彆攜帶c-myc野生型及T58A突變型基因的慢病毒載體,併包裝慢病毒,病毒滴度測定結果 分彆為6.10 × 107、5.65×107 TU/ml.結論 成功構建含有c-myc野生型及T58A突變型基因的慢病毒載體併包裝齣具高效感染力的慢病毒顆粒.
목적 구건휴대c-myc기인야생형급T58A돌변형적만병독재체.방법 응용취합매련반응(PCR)방법 확증c-myc야생형급T58A돌변형기인,이용Gateway기술구건휴대록색형광단백(GFP)기인적만병독재체,경PCR급기인측서감정후,전염293FT포장세포,산생상응만병독,측정기적도.결과 PCR화측서증실,구건분별휴대c-myc야생형급T58A돌변형기인적만병독재체,병포장만병독,병독적도측정결과 분별위6.10 × 107、5.65×107 TU/ml.결론 성공구건함유c-myc야생형급T58A돌변형기인적만병독재체병포장출구고효감염력적만병독과립.
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.
