| 微小牛蜱Bm86基因的原核表达与表达条件的优化 |
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| 引用本文： | 马米玲,关贵全,李有全,刘爱红,任巧云,牛庆丽,殷宏,罗建勋. 微小牛蜱Bm86基因的原核表达与表达条件的优化[J]. 中国寄生虫学与寄生虫病杂志, 2009, 27(6): 531-533 |
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| 作者姓名： | 马米玲 关贵全 李有全 刘爱红 任巧云 牛庆丽 殷宏 罗建勋 |
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| 作者单位： | 家畜疫病病原生物国家重点实验室,甘肃省动物寄生虫病重点实验室,中国农业科学院兰州兽医研究所,兰州 730046 |
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| 摘 要： | 根据微小牛蜱Bm86基因序列设计引物,在重组质粒pMD18-T-Bm86中克隆,并将其定向亚克隆入原核表达载体pGEX-4T-1,转化至大肠埃希菌BL21（DE3）株,用不同浓度异丙基-β-D-硫代半乳糖苷（IPTG）在不同时间进行诱导表达。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）检测结果表明,37 ℃条件下经1 mmol/L IPTG诱导8 h后,目的重组蛋白表达量最大,表达相对分子质量（Mr）约为94 000的包涵体蛋白,与预期大小一致,目的蛋白约占蛋白总量的29%。蛋白质印迹（Western blotting）分析表明,该重组蛋白可被兔抗微小牛蜱全蜱阳性血清所识别。
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| 关 键 词： | 微小牛蜱 Bm86基因 表达 |
Prokaryotic Expression of Bm86 Gene of Boophilus microplus and Optimization of the Expression Condition |
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| MA Mi-ling,GUAN Gui-quan,LI You-quan,LIU Ai-hong,REN Qiao-yun,NIU Qing-li,YIN Hong,LUO Jian-xun. Prokaryotic Expression of Bm86 Gene of Boophilus microplus and Optimization of the Expression Condition[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2009, 27(6): 531-533 |
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| Authors: | MA Mi-ling GUAN Gui-quan LI You-quan LIU Ai-hong REN Qiao-yun NIU Qing-li YIN Hong LUO Jian-xun |
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| Affiliation: | State Key Laboratory of Veterinary Etiological Biology,Key Laboratory of Veterinary Parasitology of Gansu Province,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China |
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| Abstract: | A pair of specific primers was designed based on the reported Bm86 gene of Boophilus microplus,the Bm86 gene was cloned by PCR using the plasmid pMD18-T-Bm86 as templates,and subcloned into the prokaryotic plasmid pGEX-4T-1. The recombined plasmid was transformed into E. coli BL21（DE3） and followed by expression of the protein induced by different concentration of IPTG for different time. SDS-PAGE showed that the recombinant plasmid pGEX-4T-1/Bm86 expressed a fusion protein Bm86-GST （Mr 94 000） after being induced with IPTG. High level expression of Bm86-GST was found at 1 mmol/L IPTG condition after incubation for 8 h at 37 ℃, and the expression level of the recombinant Bm86-GST reached up to 29% of total E. coli proteins. Western blotting analysis showed that the recombinant Bm86-GST was recognized by the rabbit anti-B. microplus positive serum． |
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| Keywords: | Boophilus microplus')" >Boophilus microplus Bm86 gene ')" >Expression |
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