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猪瘟病毒NS3蛋白ATPase/RNA解旋酶功能区的表达与纯化 被引量：3

Cloning Expression,Purification of Partial ATPase/RNA Helicase Functional Domain of NS3 Genes of Classical Swine Fever Virus
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摘要 采用RT-PCR技术扩增出猪瘟病毒石门株和C株NS3基因的ATPase/RNA解旋酶功能区,将其克隆到表达载体pET-28a(+)中,获得重组质粒pET-NS3SM和pET-NS3C;PCR、酶切鉴定和序列分析结果表明目的基因插入位置、方向和读码框完全正确;0.8 mmol/L IPTG诱导得到分子量为54kD的目的蛋白,Western blotting结果表明,表达的目的蛋白与猪瘟高免血清没有出现肉眼可见的反应;通过ELISA试验结果表明,重组蛋白能被CSFV阳性血清识别。 Partial ATPase/RNA helicase functional domain of NS3 genes of CSFV SHIMEN rain and C rain were amplified by RT-PCR;The recombinant plasmids pET-NS3 SMand p ET-NS3C were obtained after being cloned them into expression vector pET-28a（＋） ; And then the 54 kD target proteins were produced by inducing with 0.8 mmol/L IPTG;Western blotting showed that the expressed proteins couldn＇t be recognized by superserum against esfv. And ELISA showed that the expressed proteins could be recognized by superserum against esfv.
作者 郑杰 宁宜宝
机构地区 中国农业大学动物医学院国家动物海绵状脑病实验室 中国兽医药品监察所
出处 《中国畜牧兽医》 CAS 2007年第10期43-46,共4页 China Animal Husbandry & Veterinary Medicine
关键词 猪瘟病毒 NS3基因 ATPase/RNA解旋酶 克隆 表达 classical swine fever virus （CSFV） NS3 gene ATPase/RNA heliease cloning expression
分类号 S852.651 [农业科学—基础兽医学]
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参考文献17

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同被引文献63

  • 1刘思国,马刚,余兴龙,张茂林,涂长春.猪瘟病毒E2糖蛋白抗原表位的预测、多肽合成及特性研究[J].中国免疫学杂志,2004,20(7):448-450. 被引量：10
  • 2仇华吉,童光志,沈荣显.猪瘟兔化弱毒疫苗——半个世纪的回顾[J].中国农业科学,2005,38(8):1675-1685. 被引量：88
  • 3李国新,李娜,仇华吉,朱庆虎,李艳,王明杰,李继昌,童光志.猪瘟病毒石门株基因组全长cDNA的克隆与序列分析[J].中国预防兽医学报,2006,28(3):275-278. 被引量：11
  • 4鲁絮,张彦明,许信刚.猪瘟病毒石门株NS3基因在大肠杆菌中的表达[J].西北农林科技大学学报（自然科学版）,2007,35(2):15-18. 被引量：4
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  • 9Ganges L,Barrera M,Nunez J I,et al. A DNA vaccine expressing the E2 protein of classical swine fever virus elicits T cell respon ses that can prime for rapid antibody production and confer total protection upon viral challenge[J]. Vaccine, 2005,23 (28) : 3741 -3752.
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中国畜牧兽医
2007年 第10期

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