CAJ | 학술논문

为开展核型多角体病毒与宿主互作关系的研究,建立蓖麻蚕(Philosamia cynthia ricini)细胞体外扩增体系,分析其对不同核型多角体病毒的敏感性差异.采用剪切分离后的蓖麻蚕蛹精巢组织材料,将分散的细胞在TNM-FH培养基中经数月换液培养后,获得了扩增的蓖麻蚕蛹精巢细胞系(BMC1).BMC1的倍增时间约为65 h.病毒的感染性实验表明:BMC1细胞对苜蓿银纹夜蛾核型多角体病毒(AcNPV)比较敏感,在感染的细胞中可以明显地观察到多角体;家蚕核型多角体病毒(BmNPV)的多角体蛋白基因启动子可以在BMC1细胞中转录,但难以观察到多角体形成和细胞的病变,提示BMC1可以作为BmNPV的稳定化表达受体;而蓖麻蚕核型多角体病毒(PcrNPV)单独感染BMC1细胞时,无明显病变,只有在与AcNPV混合感染时可观察到PcrNPV多角体形成.BMC1细胞对不同核型多角体病毒的敏感性具有差异,不同核型多角体病毒的感染性差异的机制有待深入解析.
위개전핵형다각체병독여숙주호작관계적연구,건립비마잠(Philosamia cynthia ricini)세포체외확증체계,분석기대불동핵형다각체병독적민감성차이.채용전절분리후적비마잠용정소조직재료,장분산적세포재TNM-FH배양기중경수월환액배양후,획득료확증적비마잠용정소세포계(BMC1).BMC1적배증시간약위65 h.병독적감염성실험표명:BMC1세포대목숙은문야아핵형다각체병독(AcNPV)비교민감,재감염적세포중가이명현지관찰도다각체;가잠핵형다각체병독(BmNPV)적다각체단백기인계동자가이재BMC1세포중전록,단난이관찰도다각체형성화세포적병변,제시BMC1가이작위BmNPV적은정화표체수체;이비마잠핵형다각체병독(PcrNPV)단독감염BMC1세포시,무명현병변,지유재여AcNPV혼합감염시가관찰도PcrNPV다각체형성.BMC1세포대불동핵형다각체병독적민감성구유차이,불동핵형다각체병독적감염성차이적궤제유대심입해석.
To study the interactive relationship between nucleopolyhedroviruses (NPVs) and their hosts,the in vitro pro-liferation cell lines from Philosamia cynthia ricini pupal spermary tissue were established and their susceptibility to different nucleopolyhedroviruses were analyzed.By splitting the spermary tissue of Philosamia cynthia ricini pupa,the separated cells were cultured in TNM-FH medium.After several months of culture,the cell line(named as BMC1)turned into the stage of proliferation.The population doubling time of BMC1 was about 65 hours.The results of viral infection test indica-1ed lhat BMC1 cells were fairly sensitive to Autographa colifomica NPV(AcNPV).The polyhedra could be observed in the infected BMC1 cells easily.The promoter of polyhedrin gene of Bombyx mori NPV(BmNPV)could be expressed in BMC1 cells.However.formation of the polyhedra could not be found,and the infection symptoms were not observed as well.suggesting that BMC1 cells might be used for stableexpression of BmNPV.Philosamia cynthia ricini NPV (PcrNPV)alone could not cause obvious infection symp-toms.Only a mixed infection with AcNPV could lead to the expression of late polyhedrin gene and formation of PcrNPV polyhedra.The mechanism that results in the difference of susceptibility of BMC1 cells to nucleopolyhe-droviruses infection remains for further studies.