摘要
目的 :构建携HBX基因的腺病毒载体 ,为进一步研究HBX基因的功能 ,为阐明HBV感染导致肝癌的机制研究建立基础。方法 :采用PCR技术从HBV全基因组DNA中扩增HBX基因 ;将HBX基因亚克隆至质粒 pHAHA ,构建质粒 pHAHA-HBX ,酶切pHAHA -HBX质粒 ,将HAHA -HBX片段克隆到腺病毒载体 pAd -Track -CMV中 ,得到 pAdTrack -CMV -HBX ,同时与 pAdEasy - 1共转染 2 93细胞 ,确定转染效率。检测培养上清病毒中HBX的存在。结果 :将PCR扩增HBX片段成功克隆到腺病毒载体中 ,并经序列分析证实 ;建立了产病毒细胞株 ,证实重组病毒中含有HBX基因。结论
Objective:To establish a recombinant adenovirus vector carrying HBX gene for investigation of the functions of HBX gene on the hepatocellular carcinoma.Methods:HBX gene was amplified from genomic DNA of HBV by polymerase chain reaction(PCR) technique, and subcloned to plasmid pHAHA to construct recombinant plasmid pHAHA-HBX.subcloned HAHA-HBX into a shuttle vector pAdTrack-CMV.After being identified by endonuclease,and subsequently cotransformed into E.coli.293 cells with an adenoviral backbone plasmid,e.g.pAdEasy-1.Results:HBX gene was cloned from HBV genome by PCR successfully,and proved by sequence determination.A vector producing cell line 293/HBX was established.An HBX gene integration was detected by PCR in the recombinant adenovirus vectors.Conclusion:A recombinant adenovirus vector containing HBX gene was successfully constructed.
出处
《重庆医科大学学报》
CAS
CSCD
2004年第3期273-275,278,共4页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目( 3 0 3 712 74)
