减蛋综合征病毒33K蛋白基因克隆及结构分析

Cloning and Structure Analysis of 33K Protein Gene of Egg Drop Syndrome Virus

纯化的减蛋综合征病毒（ＥＤＳＶ）ＤＮＡ经ＨｉｎｄⅢ水解、０．８％琼脂糖电泳后，回收各ＤＮＡ条带并克隆至ｐＢｌｕｅｓｃｒｉｐｔＫＳ载体，建立了ＥＤＳＶ基因文库，对３３Ｋ蛋白基因进行了分析。本研究证实，ＥＤＳＶ３３Ｋ蛋白基因含有２个外显子，与羊腺病毒（ＯＡＶ）３３Ｋ蛋白比较，Ｎ端编码产物的同源性为３０．８％，Ｃ端的为６０％。用ＲＴ－ＰＣＲ扩增３３ＫｍＲＮＡ和ｃＤＮＡ克隆及序列分析证实，ＥＤＳＶ３３Ｋ蛋白含有８２碱基（ｎｔ）的内含子，去掉内含子３３Ｋ蛋白基因能形成完整的ＯＲＦ，其编码产物由１７７氨基酸（ａａ）组成，推测分子质量为２．０６×１０４，与人５型腺病毒和ＯＡＶ的同源性分别为２８．１％和４４．７％。

Egg drop syndrome virus(EDSV), AA2 strain isolated in China, was purified and the viral DNA was digested by restriction enzyme Hind . The fragments were cloned into the sites of pBluescript KS(+) vector respectively. The whole genome library was constructed and the sequence of Hind E fragment was determined. The fragment is 3 209 nt in length, encoding C terminal of 100K, 33K, p VIII and Nterminal fiber protein. The EDSV 33K protein gene is interrupted by an intron. The homology of the Nterminal and Cterminal exon of 33K with that of ovine adenovirus (OAV) is 30.8% and 60% respectively. The cDNA sequence analysis shows the 33K intron is 82 nt in length. The predicted 33K protein consists of 177aa with 2.06104 in molecular weight. The amino acid sequence has a homology of 28.1% and 44.7% to that of Ad5 and OAV respectively.