鸡bcl-2表达质粒的构建及其对转染细胞凋亡的影响  

Construction of Expression Plasmid of Chicken bcl 2 and Its Effect on Apoptosis After Transfection

作　　者：张书霞[1] 步志高 陈万芳[1] 

机构地区：[1]南京农业大学动物医学院,南京210095 [2]哈尔滨兽医研究所

出　　处：《中国农业科学》2000年第6期88-94,共7页Scientia Agricultura Sinica

基　　金：国家自然科学基金资助项目! ( No.39670 565)

摘　　要：构建了 CMV驱动及牛生长激素 poly A加尾信号修饰的 bcl- 2 c DNA真核表达质粒p CDCB1。随后 ,采用脂质体转染方法 ,将扩增并纯化的 p CDCB1转染至培养的 SPF鸡胚次代单层成纤维细胞 (CEF)内 ,96h后收集全部贴壁及上清脱落细胞 ,用 Dotblot检测 bcl- 2的表达 ,用流式细胞术测定其细胞凋亡率。结果 ,p CDCB1转染后 96h,bcl- 2在 CEF中呈高度表达 ;CEF平均凋亡率为 1 .1 4% ,明显低于转染空白载体质粒的对照组 7.64%的凋亡率 ,DNA合成期 (S期 )细胞为 2 1 .58% ,也低于对照组。结果证实鸡 bcl- 2 的表达可显著抑制体外培养 CEF凋亡进程 ,其效应与细胞 DNA合成和增殖无关。The eukaryotic expression plasmid pCDCB 1 of chicken bcl 2 that prompted by CMV and decorated by BgHpA signal was constructed. Then, the plasmid was transfected to SPF chicken embryo fibroblast (CEF) by lipofectin reagent. All cells were collected including both attached and afloat cells at 96h of incubation, and the expression of bcl 2 was determined by using dot blotting and apopotosis rate was determined by using flow cytometry.Results shown that bcl 2 expression is high in CEF and the average apoptosis rate of CEF is 1.14% after transfection pCDCB 1, the percentage is much lower compared with control group transfected vector plasmid(P<0.01). The cells of DNA synthetic phase (s phase) is 21.58%,which is lower than that of transfected vector plasmid.The finding indicated that expression of chicken bcl 2 can remarkblely suppress CEF apoptosis in vitro culture, but this effect is not correlated with DNA synthetic and cell proliferation.

关 键 词：BCL-2 真核表达质粒 转染 细胞凋亡 鸡 

分 类 号：Q78[生物学—分子生物学]