一株无血凝性兔出血症病毒的分离鉴定及其衣壳蛋白VP60基因的克隆和序列分析

畜牧兽医学报 ›› 2006, Vol. 37 ›› Issue (10): 1078-1083.doi:

一株无血凝性兔出血症病毒的分离鉴定及其衣壳蛋白VP60基因的克隆和序列分析

田浪;王红宁;李建文;廖娟;张夏兰;周万蓉;王鸿宾

1.川农业大学动物预防医学生物工程实验室，雅安 625014；2.川大学生命科学学院，成都 610064

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-10-25 发布日期:2006-10-25

Isolation and Identification of a Nonhaemagglutinating Strain of RHDV from China and Cloning and Sequence Analysis of VP60 Gene of the Strain

TIAN Lang;WANG Hong-ning;LI Jian-wen;LIAO Juan;ZHANG Xia-lan;ZHOU Wan-rong;WANG Hong-bin

1.ollege of Animal Science and Technology，Sichuan Agricultural University，Yaan 625014，China；2.ollege of Life Scince, Sichuan University, Chengdu 610064, China

Received:1900-01-01 Revised:1900-01-01 Online:2006-10-25 Published:2006-10-25

摘要/Abstract

摘要： 从临床病料中分离鉴定出1株无血凝性的致病性兔出血症病毒(RHDV)，命名为Yaan-1。HA试验测得该毒株原代及经兔体传3代后的肝毒在25 ℃、37 ℃和4 ℃时均无血凝性；但在琼脂扩散试验和对流免疫电泳中均可见分离株与常规RHDV高免血清形成清晰的沉淀线；电镜观察可见30 nm左右的病毒粒子。采用RT-PCR方法将Yaan-1株的衣壳蛋白VP60基因进行了克隆测序，序列分析表明VP60基因序列全长1 740 bp，编码579个氨基酸，测序结果提交GenBank（注册号为DQ069280），并与世界各国的常规RHDV分离株进行比较，结果表明核苷酸同源性为90.0%～98.0%，氨基酸同源性为94.1%～99.0%，氨基酸变异多发生在衣壳蛋白C、E区；同时对Yaan-1株的分子量、等电点、疏水性、跨膜区等分子结构特征进行了分析。本研究首次报道了无血凝性RHDV中国分离株，丰富了地方毒株资源，为明确我国地方株的分子流行病学，以及对VP60蛋白分子特征研究提供了新材料。

Abstract: A variant strain of rabbit hemorrhagic disease virus which was named “Yaan-1” was isolated and identified in China The virus lacked HA activity at 25,37 and 4 ℃,but gave positive results in agar diffusion reaction(ADR) and counter immunoelectrophoresis (CIE) The capsid protein（VP60）of Yaan1 strain was cloned by RT-PCR and sequenced VP60 gene of Yaan-1 was 1 740 nt in size and encoding 579 aa Alignment with other 14 sequences of RHDV isolates in the world showed that the homology were 90.0%-98.0% for nucleotide sequence，and 94.1%-99.0%for amino acid sequence respectively The results indicated that the sequence of VP60 gene of different RHDV isolates, including nonhaemagglutinating Yaan-1 strain, were highly homologous The hydrophilicity plot, flexible regions, antigenic index of VP60 protein of Yaan-1 strain, etc were also analyzed

田浪;王红宁;李建文;廖娟;张夏兰;周万蓉;王鸿宾. 一株无血凝性兔出血症病毒的分离鉴定及其衣壳蛋白VP60基因的克隆和序列分析[J]. 畜牧兽医学报, 2006, 37(10): 1078-1083.

TIAN Lang;WANG Hong-ning;LI Jian-wen;LIAO Juan;ZHANG Xia-lan;ZHOU Wan-rong;WANG Hong-bin. Isolation and Identification of a Nonhaemagglutinating Strain of RHDV from China and Cloning and Sequence Analysis of VP60 Gene of the Strain[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2006, 37(10): 1078-1083.

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