摘要
将０．９１ｋｂ的含有完整开放阅读框的人类ｂｃｌ－２ｃＤＮＡ以正向克隆于逆转录病毒载体ｐＬＸＳＮ的ＥｃｏＲⅠ位点，经ＰＡ３１７细胞包装后感染ＮＩＨ／３Ｔ３细胞，筛选出Ｇ４１８抗性克隆，经免疫印迹证实人类ｂｃｌ－２ｃＤＮＡ在ＮＩＨ／３Ｔ３细胞中获得了表达。这一表达人类Ｂｃｌ－２蛋白的哺乳类细胞模型的建立为探讨ｂｃｌ－２的作用机制及其与其它基因间关系的研究奠定了基础。
A 0.91kb EcoRⅠ insert of human bcl 2 cDNA containing the full length open reading frame was cloned in sense orientation into the EcoRⅠ site of the pLXSN retrovirus vector. The pLXSN/s bcl 2 recombinant expression vector was packaged with PA317 cells and the supernant containing high titer recombinant retroviruses was collected. NIH/3T3 cells were incubated with the supernant and the infected cells were selected for resistance to G418. Western blotting analysis showed that the G418 resistant cells could express human Bcl 2 protein. The establishment of the mammalian cell model expressing human Bcl 2 paves a way for the studies both on the mechanisms of bcl 2 action and on the interaction of bcl 2 and other genes.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1997年第4期227-229,共3页
Immunological Journal
