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杆状病毒p35基因延缓昆虫细胞凋亡

BACULOVIRUS p35 GENE DELAYS APOPTSIS IN INSECT CELLS

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【作者】 李小峰齐义鹏林宏Mallam Nock Joshua李燕李志达

【Author】 Li Xiaofeng, Qi Yipeng, Lin Hong,Mallam Nockjoshua, Li yan, Li Zhida (College of Life Sciences, Wuhan University, Wuhan 430072, China)

【机构】 武汉大学生命科学学院

【摘要】 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.

【Abstract】 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.

【关键词】 抗凋亡昆虫细胞p35基因杆状病毒
【Key words】 apoptosis inhibitinginsect cellsp35 genebaculovirus
【基金】 国家自然科学基金
  • 【文献出处】 武汉大学学报(自然科学版) ,WUHAN UNIVERSITY JOURNAL(NATURAL SCIENCE EDITION) , 编辑部邮箱 ,1998年04期
  • 【分类号】Q939.44,Q969.05
  • 【被引频次】6
  • 【下载频次】121
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