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.
出处
《生物工程学报》
CAS
CSCD
北大核心
2001年第4期445-448,共4页
Chinese Journal of Biotechnology
