蓖麻蚕核型多角体病毒DNA转染四种昆虫细胞系的研究

用蓖麻蚕核型多角体病毒(PcrNPV)DNA转染草地贪夜蛾(SF)、家蚕(Bm)、斜纹夜蛾(SL)和菜粉蝶(Pr)四种昆虫细胞系,结果表明,PcrNPV DNA能在SF细胞内复制增殖并形成多角体,Pr细胞系对PcrNPV DNA转染不敏感;Bm和SL细胞出现病变症状,但在电镜下未观察到病毒粒子或多角体。

Studies on Transfection of Four Insect Cell lines with DNA of Philosamia Cynthia ricini Nuclear Polyhedrosis Virus

The study on transfection of four insect cell lines which involve Spodoptera liturs(SL), Bombyx mori(Bm), Spodoptera frugiperda(SF)and Pieris rapae (pr)with DNA of Philosamia cynthia ricini Nuclear Polyhedrosis Virus (PcrNPV)by means of calcium phosphate coprecipitation method showed: (a)the polyhedral, particles in SF and Bm cell lines transfected with DNA of perNPV were observed under a light microscope, and no polyhedra appeared in the nuclei of SL and Pr cell lines transfected with PcrNPV DNA. (b)four insect cell lines were detected with an electron microscope, the polyhedra and virions of PcrNPV were only observed in SF cell line transfected with PcrNPV DNA, and whether polyhedrin or virion did not appear in the other three cell lines transfected with DNA of PcrNPV as well as four control groups. (c)total DNA of both the transfected SF cell line and the control group was repectively cleaved with BamHI and EcoRI, the viral DNA bands were only observed in the DNA sample of the transfected SF cell line and the restriction pattern of the viral DNA from the transfected SF cell line was identical to the donor PcrNPV DNA. The result of the experiment showed PcrNPV DNA can not only transfect SF cell line but also reproduce and build polyhedra in the nuclei of SF cell. Therefore, it was proposed that SF cell line should be used as a receptor system which is useful for study on molecular biology and genetics of PcrNPV DNA and expression of extraneous genes carried in the vector of PcrNPV.