摘要
根据已发表的猪瘟病毒序列 ,设计并合成了覆盖有 NS2 - 3全基因的 4对引物 ,应用RT -PCR技术从接种了猪瘟兔化弱毒 (hogcholeraviruslapinizedChinesestrain ,HCLV)的兔脾组织中 ,成功地扩增了 NS 2 - 3基因 ,将其克隆到T载体后 ,测定了其核苷酸序列。结果显示 ,所克隆的NS 2 - 3基因长 2 96 4个核苷酸 ,编码 988个氨基酸。用DNASIS和PROSIS软件分析表明 ,该基因具有丝氨酸类蛋白酶活性及含有解旋酶超家族特征性保守序列。核苷酸序列同源性分析表明 ,HCLV与国外报导的 8株HCV毒株“C”、Riems、ALD、GPE、Glentorf、CAP、Brescia和Alfort株NS 2 - 3基因核苷酸序列同源性分别为 99%、99%、96 %、96 %、95 %、95 %、94%和 86 % ,推导的氨基酸序列同源性分别为 99%、99%、98%、98%、98%、97%、97%和 95 % ,氨基酸序列的高度同源性表明了NS 2 - 3是猪瘟病毒中高度保守的蛋白质之一 ,这与该基因产物参与病毒复制及多聚蛋白前体加工过程中的重要作用相对应。
According to the published nucleotide sequence of genome of hog cholera virus(HCV), 4 pairs of specific primers were designed and synthesized. From the spleen of rabbits infected with HCLV, the NS2-3 gene was amplified by RT-PCR. The amplified fragments were cloned and sequenced. The results showed that the NS2-3 gene sequenced was 2,964 base pair long which encoded 988 amino acids and the nonstructural protein NS2-3 was to be both a serine-type protease and a nucleotide triphosphatase (NTPase)/RNA helicase. The nucleotide sequence homologies of NS2-3 gene of HCLV strain with the 'C', Riems, ALD, GPE -, Glentorf, CAP, Brescia and Alfort strains were 99%, 99%, 96%, 96%, 95%, 95%, 94% and 86% respectively and the amino acid sequence homologies were 99%, 99%, 98%, 98%, 98%, 97%, 97% and 95% respectively. It revealed that the NS2-3 protein was one of the most conserved viral protein among HCV strains.
出处
《病毒学报》
CAS
CSCD
北大核心
2000年第2期150-154,共5页
Chinese Journal of Virology
基金
国家攀登B计划!( 85 -4 1子专题 )
关键词
中国猪瘟兔化弱毒
NS2-3基因
序列分析
疫苗
hog cholera virus lapinized Chinese strain (HCLV)
NS2-3 gene
sequence analysis
