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猪肺炎霉形体Dnak-P97融合基因的表达
及其产物的生物学活性
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王亮1，乔祖健1，2，李媛1，张建华1，3，梁立1，4，辛九庆1*
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（1.中国农业科学院 哈尔滨兽医研究所，黑龙江 哈尔滨150001；2.东北农业大学，黑龙江 哈尔滨150030；
3.北京英博教育咨询中心，北京100086；4.黑龙江畜牧兽医职业学院，黑龙江 双城150111）
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摘要：
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应用DNA重组技术，将猪肺炎霉形体黏附因子P97 C末端基因与猪肺炎霉形体伴侣蛋白Dnak C末端基因同时克隆到pET-30a表达载体上，构建了重组表达载体。将鉴定正确的重组质粒转化大肠杆菌BL21，用终浓度为1.0mmol/L的IPTG诱导6h。用BugBusterTM Ni-NTA His·Bind纯化试剂盒在自然条件下对目的蛋白进行了纯化；经Western-blotting和动物试验，证实，该蛋白具有良好的生物学活性。 |
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关键词：
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猪肺炎霉形体；猪霉形体肺炎；P97基因；伴侣蛋白Dnak；热休克蛋白70 |
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中图分类号：S 852.62：Q 786
文献标识码：
A 文章编号：1673-4696(2008)02-0123-05 |
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Expression of Dnak-P97 fusion gene of Mycoplasma hyopneumoniae
and biological activities of the expressed product |
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WANG Liang1,QIAO Zu-jian1,2，LI
Yuan1，ZHANG Jian-hua1,3,LIANG Li1,4,XIN
Jiu-qing1 |
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(1.Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China；2.Northeast
Agricultural University,Harbin 150030,China 3.Beijing U-Bridge Education Consulting Center,Beijing 100086,China；4.Heilongjiang Vocational College of Animal Husbandry and Veterinary Science,Shuangcheng 150111,China) |
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Abstract: The C-terminal Mof P97 gene and C-terminal Dnak gene of Mycoplasma hyopneumoniae(Mhp) were linked and co-cloned into pET-30a expression vector by DNA recombination technique to construct recombinant vector P97-Dnak-pET-3a.The positive recombinant plasmids were transformed into host strain Escherichia coli BL21 and induced with 1.0mmol/L IPTG for 6 hours.The expressed protein was purified with BugBusterTM Ni-NTA His·Bind purification kit and its biological activities were identified by Western-blotting and experimental infection of animals. |
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Key words: Mycoplasma hyopneumoniae;mycoplasmal pneumonia of swine;P97 gene；chaperone Dnak；heat shock protein 70 |
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