实用癌症杂志
實用癌癥雜誌
실용암증잡지
The Practical Journal of Cancer
2015年
12期
1755-1757
,共3页
邱宇安%陈文学%靳文剑%陈火国%钟宁%余洁丽
邱宇安%陳文學%靳文劍%陳火國%鐘寧%餘潔麗
구우안%진문학%근문검%진화국%종저%여길려
LeftyA%慢病毒载体
LeftyA%慢病毒載體
LeftyA%만병독재체
LeftyA%Lentiviral vector
目的:想构建LeftyA基因的慢病毒表达载体,鉴定其在293T细胞中的表达。方法首先扩增LeftyA基因序列,将其与慢病毒骨架载体连接后,鉴定慢病毒表达载体构建成功。将重组慢病毒表达载体质粒及包装质粒共转染293T细胞,鉴定慢病毒转染并包装成功。结果 LeftyA基因成功转导至以GV287为骨架质粒的慢病毒系统中,包装后测定病毒滴度达2.0×108 TU/ml,免疫印记法检测到LeftyA蛋白在293T细胞中的表达。结论成功构建了携带人LeftyA基因的慢病毒表达系统。
目的:想構建LeftyA基因的慢病毒錶達載體,鑒定其在293T細胞中的錶達。方法首先擴增LeftyA基因序列,將其與慢病毒骨架載體連接後,鑒定慢病毒錶達載體構建成功。將重組慢病毒錶達載體質粒及包裝質粒共轉染293T細胞,鑒定慢病毒轉染併包裝成功。結果 LeftyA基因成功轉導至以GV287為骨架質粒的慢病毒繫統中,包裝後測定病毒滴度達2.0×108 TU/ml,免疫印記法檢測到LeftyA蛋白在293T細胞中的錶達。結論成功構建瞭攜帶人LeftyA基因的慢病毒錶達繫統。
목적:상구건LeftyA기인적만병독표체재체,감정기재293T세포중적표체。방법수선확증LeftyA기인서렬,장기여만병독골가재체련접후,감정만병독표체재체구건성공。장중조만병독표체재체질립급포장질립공전염293T세포,감정만병독전염병포장성공。결과 LeftyA기인성공전도지이GV287위골가질립적만병독계통중,포장후측정병독적도체2.0×108 TU/ml,면역인기법검측도LeftyA단백재293T세포중적표체。결론성공구건료휴대인LeftyA기인적만병독표체계통。
Objective To construct LeftyA gene lentiviral vector system and determine its expression in 293Tcells. Methods Full-length sequence of LeftyA was amplified by PCR,and was connected with lentiviral vector.Gene sequencing was used to identify that recombinant lentiviral vector was constructed successfully.The plasmids were transfected into 293Tcells.In-dentification by fluorescent microscopy confirmed lentiviral transfection and packaging successfully.Results LeftyA were sub-cloned into GV287 as skeleton plasmid for a lentivirus packing system.The virus titer was 2.0 ×108 TU/ml in the supernatant liq-uid.The proteins of LeftyA were detected by western blot.Conclusion The LeftyA gene recombinant lentiviral expression system is constructed successfully.