猪瘟病毒石门株与兔化弱毒株E0糖蛋白基因的克隆及序列分析

CLONING AND SEQUENCE ANALYSIS OF E0 GENE OF HOG CHOLERA VIRUS LAPINIZED CHINESE STRAIN AND VIRULENT SHIMEN STRAIN

参考已发表的猪瘟病毒序列，设计并合成了一对引物，应用ＲＴＰＣＲ 技术，扩增了猪瘟兔化弱毒（Ｈｏｇ ｃｈｏｌｅｒａ ｖｉｒｕｓ ｌａｐｉｎｉｚｅｄ Ｃｈｉｎｅｓｅ ｓｔｒａｉｎ ， ＨＣＬＶ） 和石门强毒株的Ｅ０ 糖蛋白基因，并将其克隆到ｐＧＥＭＴ 载体中，测定了其核苷酸序列，并推导了其氨基酸序列。结果表明我国这两株强弱不同毒株Ｅ０ 糖蛋白核苷酸序列同源性和推导的氨基酸序列同源性分别为９５－０ ％ 和９４－３ ％ ，有１３ 个氨基酸的差异，ＨＣＬＶ 比石门株多了一个潜在的Ｎ糖基化位点。将我国这两株病毒与国外已报导的ＨＣＶ 毒株Ｅ０ 基因序列进行了比较，发现石门株与日本的两株毒株ＡＬＤ 和ＧＰＥ－ 同源性较高，核苷酸序列同源性分别为９７－４ ％ 和９６－５ ％ ，氨基酸同源性分别为９７－４ ％ 和９６－０ ％ ，而与欧洲Ｂｒｅｓｃｉａ 株和Ａｌｆｏｒｔ 株同源性较低，核苷酸同源性分别为９２－２ ％ 和８６－５ ％ ，氨基酸同源性为９５－２ ％ 和９２－５ ％ ， ＨＣＬＶ 与ＡＬＤ、ＧＰＥ－ 、Ｂｒｅｓｃｉａ、Ａｌｆｏｒｔ 株核苷酸同源性分别为９５－６ ％ 、９４－９ ％ 、９１－３ ％ 、８５－５ ％ ，推导的氨基酸同源性分别为９３－４ ％ 、９２－５ ％

According to the published nucleotide sequences of genome of hog cholera virus, one pair of specific primers were designed and synthesized. From the spleen of rabbits which were infected with HCLV and HCV Shimen strain infected pig blood, the two E0 genes were amplifed by RT\|PCR. The amplified fragments were cloned into pGEM\|T vector and sequenced. Sequence analysis showed nucleotide sequence and deduced amino acid sequence homologies of the E0 gene between HCLV and Shimen strain were 95.0% and 94.3%. There was 13 amino acid substitutions between them. One N\|glycosylation site was missing from E0 gene of the Shimen strain. The nucleotide sequence homologies of the Shimen strain with the ALD, GPE\+-, Brescia and Alfort strains were 97 4%, 96.5%, 92.2% and 86.5%, respectively, the deduced amino acid sequence homologies were 97.4%, 96.0%, 95.2% and 92.5%, respectively. The nucleotide sequence homologies of E0 gene of the HCLV strain with the ALD, GPE\+-, Brescia and Alfort strains were 95.6%, 94 9%, 91.3% and 85.5%, respectively, the deduced amino acid sequence homologies were 93 4%, 92.5%, 91.6% and 90.7%, respectively. E0 gene from HCV was shown to be similar to a family of fungal and plant ribonuclease. The catalytically important residues were 28 to 40 and 71 to 89 of HCV E0 gene.