杆状病毒p35基因延缓昆虫细胞凋亡

杆状病毒p35基因延缓昆虫细胞凋亡 BACULOVIRUS p35 GENE DELAYS APOPTSIS IN INSECT CELLS 李小峰 Li Xiaofeng 1 first-author 齐义鹏 Qi Yipeng 1 林宏 Lin Hong 1 Ｍａｌｌａｍ Ｎｏｃｋ Ｊｏｓｈｕａ Mallam Nockjoshua 1 李燕 Li yan 1 李志达 Li Zhida 1 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences, Wuhan University College of Life Sciences, Wuhan University 构建了以新霉素抗性基因为筛选标记的带有ｐ３５基因的转移载体ｐ３５ＩＥ１Ｎｅｏ，以此载体转染Ｓｆ９细胞，经Ｇ４１８筛选得到染色体上稳定整合质粒ｐ３５ＩＥ１Ｎｅｏ的Ｓｆ９细胞，克隆化培养后取名为Ｓｆ９－３５．经细胞原位杂交实验证明，Ｓｆ９－３５细胞的染色体ＤＮＡ上含有ｐ３５基因；经放线菌素Ｄ处理后的细胞核酸电泳检测，证实Ｓｆ９－３５具有抗凋亡特性能够支持凋亡抑制基因缺失的ｖＡｃΔｐ３５病毒复制；发现ｐ３５基因在细胞内组成型表达只能延缓ｖＡｃΔｐ３５感染及放线菌素Ｄ处理诱发的细胞凋亡的过程，而不能完全阻止其诱发的细胞凋亡． We constructed an expressing vector p35IE1Neo containing antiapoptotic p35 gene and neomycin resistant gene（as a selection marker）. After transfcted Sf9 cells with p35IE1Neo and by G418 screening, we got transformed cells that appeared resistant to G418 and picked one clone named Sf9 35. By hybridization in situ , it was found that p35 gene had integrated into the chromosome of Sf9 35 cells; It was found that Sf9 35 cells surported the replication of vAcΔp35 deleted apoptosis inhibiting gene. After actinomycin D treatment and cellular DNA electrophoresis, Sf9 35 cells were found to resist apoptosis induced by infection of vAcΔp35 deleting p35 gene and actinomycin D treatment; And it was also found that apoptosis induced by viral infection and actinomycin D treatment can only be delayed, but can not be stopped in Sf9 35. 抗凋亡 昆虫细胞 p35基因 杆状病毒 apoptosis inhibiting insect cells P35 gene baculovirus Q939.44;Q969.05 国家自然科学基金(61672102) 1998-04-01 2021-04-01 4