轮状病毒1、2血清型VP7蛋白基因的扩增及其克隆  

Amplification and Cloning of Gene Encoding Outer Capsid Protein VP7 from Human Rotavirus Serotypes 1 and 2

作　　者：陈钧[1] 纪绍忠[1] 谈幸之[1] 范明远[1] 

机构地区：[1]中国预防医学科学院流行病学微生物学研究所,卫生部食品卫生监督检验所

出　　处：《中华实验和临床病毒学杂志》1994年第4期312-315,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：用聚合酶链反应扩增了轮状病毒Ｗａ株（第１血清型）和ＤＳ－１株（第２血清型）的全长ＶＰ７蛋白基因，利用两引物５＇末端的多接头位点，以ｐＵＣ１８质粒为载体构建了含两株ＲＶＶＰ７基因ｃＤＮＡ拷贝的重组体。经ＰＣＲ扩增制备得１～４血清型ＲＶＶＰ７基因高变区（５１－３９２核苷酸）￣（３２）Ｐ标记核酸探针，对重组质粒型别进行了Ｓｏｕｔｈｅｒｎ和斑点杂交鉴定，结果Ｗａ株ＶＰ７基因重组体只与１型探针呈强杂交信号，ＤＳ－１株重组体只与２型探针呈强杂交信号。The full length gene encoding outer capsid protein VP7 was amplified from human rotavirus serotypes 1(strain Wa)and 2(strain DS-1).Using the polylinker devised on the primer pair, the cDNA of VP7 gene was inserted into plasmid and the recombinant plasmid was used to transform E.coli JM 103.￣(32)p-labeled probes from hyperdivergent region (nucleotides 51 to 392) of rotavirus serotypes 1 to 4 gene encoding the VP7 glyprotein were synthesized by using polymerase chain reaction method,and were used to identify the recombinant plasmid by Southern or dot hybridization. The recombinant plasmid of strain Wa hybridized only to the probe of serotype 1 and strain DS-1 only to the probe of serotype 2.

关 键 词：轮状病毒 基因克隆 血清型 蛋白基因 

分 类 号：R373.24[医药卫生—病原生物学]