文献类型：期刊文献

中文题名：人神经营养因子-3重组逆转录病毒的包装与鉴定

英文题名：The Package and Identifying of hNT-3 Recombinated Retroviral Vector

作者：欧阳立明[1];周涛[2];薛冲[2];刘志敏[2];张嗣良[1]

机构：[1]华东理工大学生物工程学院,上海200237;[2]军事医学科学院生物工程研究所,北京100071

年份：2001

卷号：17

期号：4

起止页码：445

中文期刊名：生物工程学报

外文期刊名：Chinese Journal of Biotechnology

收录：CSTPCD;;Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；PubMed;

语种：中文

中文关键词：逆转录病毒载体;pLXNS;人神经营养因子-3;重组病毒;包装;鉴定

外文关键词：retroviral vector pLXSN, human neurotrophin-3(hNT-3), gene therapy

摘要：在逆转录病毒载体pLXSN中引入人神经营养因子 3(hNT 3) ,构建成重组质粒pLXSN NT3,转染包装细胞PA317后经G418筛选得到几个稳定产毒的细胞克隆。测定这几个克隆的病毒滴度 ,最高达到 1 6 0× 10 5 ,且产毒细胞株不产生复制型病毒。PCR和大乳鼠背根神经节活性检测证明hNT 3已整合到宿主细胞基因组并能稳定表达。
hNT-3 was inserted into retroviral vector pLXSN to get pLXSN-NT3,and it was transfected into packaging cell line PA317 to form G418 resistant cell clones.The G418 resistant clones were titered and checked for the existence of replication viral.Genome DNA isolated from the cell clone of highest titeration showed the function gene,hNT-3 cDNA,had integrated into the genome of host cells verified by PCR.And bioassay of the clone's cell culture supernant exhibited that it can induce neurite overgrowth in the primary culatures of rat spinal cord dorsal root ganglions as compared with controls.