Inhibiting Mechanism of Baculovirus p35 Gene to Apoptosis

We transfected Sf9 cells with an expressing vector p35IE1Neo containing antiapoptotic p35 gene and neomycin-resistant gene (as a selection marker). 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; By using 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.

两种昆虫杆状病毒诱导的细胞凋亡

首次发现粉纹夜蛾多粒包埋核多角体病毒（Ｔｒｉｃｈｏｐｌｕｓｉａｎｉｍｕｌｔｉｃａｐｓｉｄｎｕｃｌｅａｒｐｏｌｙ－ｈｅｄｒｏｓｉｓｖｉｒｕｓ，ＴｎＭＮＰＶ）和中国棉铃虫单粒包埋核多角体病毒（Ｈｅｌｉｏｔｈｉｓａｒｍｉｇｅｒａｓｉｎｇｌｅｃａｐ－ｓｉｄｎｕｃｌｅａｒｐｏｌｙｈｅｄｒｏｓｉｓｖｉｒｕｓ，ＨａＳＮＰＶ）分别诱导斜纹夜蛾细胞Ｓｌ－ｚｓｕ－１和粉纹夜蛾细胞Ｔｎ－５Ｂ１－４发生典型的细胞凋亡．Ｓｌ－ｚｓｕ－１和Ｔｎ－５Ｂ１－４的细胞凋亡分别开始于病毒感染后１０ｈ和２４ｈ左右，其高峰期细胞凋亡率分别为９５％～１００％和６０％左右．约１０％的Ｔｎ－５Ｂ１－４细胞产生病毒包涵体．感染复数为５的病毒可诱导２种细胞的凋亡率达到高峰．ＴｎＭＮＰＶ和ＨａＳＮＰＶ的ＤＮＡ不能诱导细胞凋亡，只有用含芽生病毒的接种液攻击细胞才能诱导细胞凋亡．

杆状病毒凋亡抑制基因的研究进展

杆状病毒(bacu lovirus)感染昆虫细胞能引起细胞凋亡,但在长期进化过程中,杆状病毒可通过自身编码凋亡抑制基因的表达,抑制细胞凋亡以利于自己的增殖。目前在杆状病毒基因组中已发现两种不同类型的细胞凋亡抑制基因p35/p49和iap,这两类凋亡抑制基因分别作用于细胞凋亡途径的不同位点,以抑制细胞的凋亡。近年来人们对这两种基因的蛋白结构及作用机制等方面进行了大量的研究,这些为今后研究昆虫细胞凋亡,扩大杆状病毒宿主范围等方面奠定了基础。