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作 者:周建华[1] 丛国正[1] 高闪电[1] 常惠芸[1]
机构地区:[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,国家口蹄疫参考实验室,甘肃兰州730046
出 处:《华北农学报》2008年第6期28-31,共4页Acta Agriculturae Boreali-Sinica
基 金:国家科技支撑计划(2006BAD06A14;2006BAD06A10)
摘 要:以口蹄疫病毒株OA/58 RNA为模板,反转录并扩增目的cDNA,然后与pMD18-T载体连接并转化JM109菌株,提取的重组质粒用凝胶电泳、PCR和BarnHⅠ、HindⅢ双酶切法鉴定。分析表明,口蹄疫病毒VP1、VP2、VP3和VP4在核苷酸水平上的变异率是无差异的(P〉0.05);而它们在氨基酸水平上的变异率差异显著(P〈0.05)。该毒株与20株源于GenBank中的VP3氨基酸序列比较发现其保守区主要位于第1~24、26~35、37~43、45~57、61~122、124~173、175~209、211~220位。运用同源模建,OA/58 VP3蛋白三维空间结构可分为A,B,C3个结构区域。保守区氨基酸残基在维持VP3蛋白的空间构象和功能方面具有重要作用。Foot-and-mouth disease virus strain OA/58 RNAs were used as templates for RT-PCR. The amplified cDNA products were cloned into pMD18-T vectors and transformed into JM109. The recombinant plasmids were identified by electrophoresis, PCR, and analysis of tow cleavages with BamH Ⅰ and Hind Ⅲ. After analyzing the difference among VP1, VP2, VP3, VP4, at the nueleotide level, the mutation rates of these 4 encoding sequenees were no difference (p 〉 0.05), however, at the amino acid level, those mutation rates were different (p 〈 0.05). The regions of 1 - 24th, 26 - 35th, 37- 43th, 45 -57th, 61 - 122th, 124- 173th, 175- 209th and 211 -220th in VP3 protein most probably are conservative. Depending on homology modeling the FMDV strain OA/58 VP3 protein, the 3D mold was gained. And this 3D mold was divided into A, B and C regions. This research should be used as an instruetion in order to direct the work on FMDV VP3 protein.
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