中国猪瘟兔化弱毒株（C-株）兔脾组织毒主要保护性抗原E2（gp55）基因序列分析

摘要

The major protective antigen E2 gene of Chinese Hog Cholera Lapinised Virus（C-strain）directly from rabbit spleen tissue，was amplified by reverse transcription （RT） and nested-polymerase chain reaction（N-PCR） and successfully cloned，it′s sequence was determined and compared with those of other virulent strains of HCV published natively or abroad．The result showed that，the nucleotide homologies of E2 gene of C-strain directly from rabbit spleen tissue with C strain（adapted to SK6 cell），C-strain（adapted to calf testicle cell，HCLV-C），HCV-SM strain（virulent strain Shimen），virulent strain Brescia（Netherlands）and virulent strain Alfort（Germany），were 98.87％，98.34％，94.58％，91.00％ and 80.78％ respectively．The amino acid homologies were respectively 98.95％，97.37％，94.22％，91.60％ and 89.23％．In three neutralized antigenic domains A、B、C of E2 gene compared with the strain described aboved and epidemic virulent strain，there was very little difference or even no difference between C-strain from rabbit spleen and C-strain adapted to cells，while the difference between C-strain form rabbit spleen and epidemic virulent strain，classic virulent strain in B、C domains was great．Morever compared with chinese classic virulent strain Shimen，there was great difference between our countrys′ virulent strains in 1980s and those in 1990s，this showed that the epidemic virulent strains of hog cholera virus have changed in our country．%利用反转录（RT）及套式PCR（N-PCR）方法扩增了中国猪瘟兔化弱毒株（C-株）兔脾组织毒主要保护性抗原E2（gp55）基因，成功地将其克隆并测定了核苷酸序列，与国内外已发表的猪瘟病毒（HCV）E2基因序列比较的结果是：C-株兔脾毒与C-株细胞（SK6）毒、C-株疫苗（犊牛睾丸细胞，HCLV-C）毒、HCV-SM株（石门）毒、Brescia株（荷兰）毒、Alfort株（德国）毒的E2核苷酸序列同源性分别为98.87％、98.34％、94.58％、91.00％、80.78％；氨基酸同源性分别为98.95％、97.37％、94.22％、91.60％、89.23％。对C-株兔脾毒与C-株细胞毒、经典强毒及国内流行野毒E2上的A、B、C三个中和性抗原区的氨基酸组成进行了比较，其结果为：C-株兔脾毒与C-株细胞毒的差异很小甚至没有差异，而与流行野毒及经典强毒在B、C区有较大的差异。我国经典强毒石门毒与国内80年代和90年代流行毒之间有明显的差异，表明我国猪瘟流行毒株发生了变化。