猫杯状病毒感染性克隆的构建  

Construction of infectious clone of Feline Calicivirus

作　　者：郑翠玲[1,2] 向华[1] 黄元[1] 陈晶[1] 王晓虎[1] 宣华[1] 

机构地区：[1]广东省农业科学院动物卫生研究所,广东广州510640 [2]唐山职业技术学院,河北唐山064003

出　　处：《中国兽医杂志》2014年第11期44-46,共3页Chinese Journal of Veterinary Medicine

基　　金：广东省自然科学基金项目(8151064001000004);广东省农业攻关项目(2007A020300005-7);广东省农业攻关重点项目(2006A20301006)

摘　　要：为了构建猫杯状病毒感染性克隆,在对FCV CH-GD株全长基因组测序基础上,引入单一酶切位点和T7 RNA聚合酶启动子,用重组PCR方法,获得全长基因组c DNA,体外转录后转染F81细胞。结果成功构建含单一酶切位点的FCV全长c DNA克隆,体外转录并转染F81细胞,培养物上清液对F81细胞有感染性,细胞出现典型病变。表明获得了FCV CH-GD株的感染性克隆,为进一步研究杯状病毒的生物学特性和致病机理及研发新的疫苗奠定了基础。Objective：To construct infectious clone of Feline Calicivirus（FCV）.Methods：Based on full-length genomic sequence of FCV CH-GD strain,the single restriction sites and T7 RNA polymerase promoter were introducted.By recombinant PCR,the full-length genomic c DNA was obtained.After RNA was transcribed in vitro,the resulted RNA was transfected into F81 cells.Results：FCV full-length c DNA clone containing a single enzyme cutting site was constructed successfully,and then was transcribed in vitro and transfected into F81 cell. The culture supernatant was infectious to F81 cells,and led to classical cytopathic effect（CPE）.Conclusion：The infectious clone of FCV CH-GD strain was obtained.It lays a foundation for further study on the biological characteristics of caliciviruse and pathogenic mechanism and development of a new vaccine.

关 键 词：猫杯状病毒 感染性克隆 RT-PCR RNA 

分 类 号：S858.293[农业科学—临床兽医学]