诊断学理论与实践2012，Vol.11Issue(3)：295-298,4.DOI:10.3969/j.issn.1671-2870.2012.03.020
三元复合因子Net嵌合型重组腺病毒载体的构建及鉴定
Construction and identification of chimeric recombinant adenovirus vector with ternary complex factor Net gene
摘要
Abstract
Objective To clone human full-length ternary complex factor Net gene and construct its recombinant adenovims vector.Methods Full length Net gene was amplified by reverse transcription polymerase chain reaction ( RT-PCR ) from human pancreatic tissues,and 293 cells were co-tranfected with adenovirus skeleton plasmid pBHG35 and shuttle plasmid pDC316- Net by homogenous recombination method,and adenovirus recombinant plasmid Ad5/ F35-Net was obtained.A high-titer adenovirus recombinant plasmid Ad5/ F35-Net was generated after packing and amplification in the 293 cell.The expression of Net gene was detected by PCR and Western blotting.Result cDNA fragment of 1224 bp was amplified by RT-PCR from human pancreatic cancer tissue,and sequencing identified it to be the Net gene.A high-titer recombinant adenovirus vector Ad5/F35-Net was constructed and packed.Conclusions Human ternary complex factor Net gene is successfully cloned and its recombinant adenovirus vector is constructed,which provides a foundation for further study on biological role of Net gene in related tumors.关键词
腺病毒载体/Net基因/克隆/293细胞Key words
Adenovirus vector/Net gene/Clone/293 cells分类
医药卫生引用本文复制引用
李百文,倪培华..三元复合因子Net嵌合型重组腺病毒载体的构建及鉴定[J].诊断学理论与实践,2012,11(3):295-298,4.基金项目
上海市自然科学基金项目11ZR1429000 （）
