J亚群禽白血病病毒Hrb-1和JL-2株囊膜基因的克隆及遗传分析

Genetic analysis of avian leukosis virus subgroup J Hrb-1 and JL-2 strain’s env gene

采用特异性检测J亚群禽白血病病毒的RT PCR方法 ,自哈尔滨市及吉林省患病鸡群中分离鉴定出 2株J亚群禽白血病病毒 ,分别命名为Hrb 1和JL 2。 2株病毒经SPF鸡胚成纤维细胞增殖 ,获取其前病毒DNA。采用依据原型毒株HPRS 10 3cDNA序列设计并合成的 1对引物 ,进行PCR扩增 ,得到了JL 2和Hrb 1的囊膜基因。分别构建了重组质粒 pMD18 T JL2 /env和pMD18 T Hrb 1/env。在对 2株病毒囊膜基因进行序列测定的基础上 ,将囊膜基因中 gp85和 gp37的核苷酸序列及其推导氨基酸序列与国际上不同的参照毒株以及国内部分分离株进行了比较分析 ,结果表明 ,JL 2和Hrb 1分别与美国病毒株adol hc 1和UD3有着较大的亲缘关系。

Two strains of ALV-J named Hrb-1 and JL-2 were identified by reverse transcription-polymerase chain reaction(RT-PCR), These viruses were propagated on the SPF CEF and harvested after 7 days culturing. According to the sequence of prototype ALV-J virus HPRS-103 cDNA gene published, a pair of primers were designed to amplify part of the env gene including gp85,gp37 and E-element by polymerase chain reaction(PCR).The pMD18-T-JL2/env和pMD18-T-Hrb-1/env were obtained by inserting the DNA fragments including env genes of two strains into pMD18-T vector. After being sequenced the genetic and antigenic analysis of Hrb-1 and JL-2 strains about gp85 and gp37 were performed on level of the amino acid and nucleic acid sequence, the result showed that Hrb-1 and JL-2 strains were related to ALV-J adol-hc-1 and UD3 strains, respectively.