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2002～2004年间从国内养殖鲤科鱼类和观赏鱼类中分离出8株鲤春血症病毒(SVCV)。根据SVCV参考株 全序列,设计引物,用逆转录聚合酶链式反应扩增出8株SVCV糖蛋白编码基因片段,并对扩增产物进行了克隆和 序列测定。用生物信息学方法对测得的序列进行分析,结果8个国内分离株的糖蛋白基因序列与参考株的基因序 列相似性均在92％以上,8个国内分离株之间基因序列相似性均在97．7％以上;8个国内分离株之间糖蛋白推导 出的氨基酸序列相似性均在94．5％以上,与参考株氨基酸序列相似性在92．9％-94．9％之间。系统发育树分析结 果表明,SVCV国内分离株与USA株、980451株、980528株和970469株的进化方向一致,与其它SVCV毒株在进 化方向上不同。8个毒株有19个共同的酶切位点,推导出的氨基酸序列中有10个亲水区、10个可能的抗原位点 和10个跨膜蛋白区域,其峰值基本一致。对SVCV国内分离株的糖蛋白6个功能位点(天冬酰胺糖基化位点、精 氨酸-甘氨酸-天冬氨酸序列、酪蛋白激酶Ⅱ磷酸化位点、蛋白激酶C磷酸化位点、酪氨酸磷酸化位点和肉豆蔻酰 基化位点)进行了初步分析。
From 2002 to 2004, 8 strains of Cyprinid carp virus (SVCV) were isolated from domestic carps and ornamental fish. According to the complete sequence of SVCV reference strain, primers were designed, and 8 SVCV glycoprotein encoding gene fragments were amplified by reverse transcription polymerase chain reaction. The amplified products were cloned and sequenced. The bioinformatics method was used to analyze the measured sequences. The results showed that the similarity of the sequence of the glycoprotein genes of 8 domestic isolates with that of the reference strains was over 92%. The similarity of the gene sequences of 8 domestic isolates Over 97.7%. The amino acid sequences deduced by the glycoproteins of 8 domestic isolates shared more than 94.5% identities with the amino acid sequences of 92.9% -94.9% . The results of phylogenetic tree analysis showed that the SVCV domestic isolates had the same evolutionary direction as the USA strains 980451, 980528 and 970469, but different from other SVCV strains in evolutionary direction. Nineteen common restriction sites were found in the eight strains, and the deduced amino acid sequence had 10 hydrophilic regions, 10 possible antigenic sites and 10 transmembrane protein regions with the same peak. Six functional sites of glycoprotein (asparagine glycosylation site, arginine-glycine-aspartic acid sequence, casein kinase Ⅱ phosphorylation site, protein kinase C phosphorylation site Point, tyrosine phosphorylation sites and myristoylation sites).