| 牛整合素β6基因的原核表达及其抗血清的制备研究 |
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| 引用本文： | 吕洋,陈莉莉,王洪梅,武建明,何洪彬,李杰.牛整合素β6基因的原核表达及其抗血清的制备研究[J].安徽农业科学,2011(9):5255-5257,5342. |
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| 作者姓名： | 吕洋 陈莉莉 王洪梅 武建明 何洪彬 李杰 |
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| 作者单位： | 东北农业大学生命科学学院;山东省农业科学院奶牛研究中心; |
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| 基金项目： | 国家转基因重大专项(2009ZX08007-006B); 国家自然科学基金项目(31072160); 山东省农业重大应用技术创新课题; 山东省科技攻关课题(2009GG20002032); 济南市高校院所自主创新计划项目(201004027); 山东省自然科学基金项目(ZR2009DQ016) |
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| 摘 要： | 目的]为深入研究ITGB6亚基的基因功能奠定基础。方法]利用基因克隆技术获得整合素β6基因（ITGB6）胞外区,然后将其重组到pET-32a（＋）质粒中,经鉴定及序列分析确定获得了重组阳性克隆;转化大肠杆菌BL21感受态细胞,IPTG诱导及蛋白纯化。结果]SDS-PAGE结果显示,在94 kD处出现特异性条带;对该融合蛋白进行分离纯化后免疫新西兰兔,制备ITGB6蛋白抗血清,Western-blot分析表明抗血清可与原核表达的ITGB6蛋白特异性结合,ELISA方法检测抗血清的效价可达到1∶2 560。结论]成功表达了牛的重组整合素β6基因。
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| 关 键 词： | 整合素β6基因 原核表达 抗血清 |
Expression of Bovine Integrin β6 Gene and Preparation of Its Antiserum |
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| LV Yang et al.Expression of Bovine Integrin β6 Gene and Preparation of Its Antiserum[J].Journal of Anhui Agricultural Sciences,2011(9):5255-5257,5342. |
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| Authors: | LV Yang |
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| Institution: | LV Yang et al(College of Life Science,Northeast Agriculture University,Haerbin,Heilongjiang 150030) |
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| Abstract: | Objective] The aim was to lay a foundation for deep studies on gene function of ITGB6 subunit.Method] The integrin β6(ITGB6) gene extracellular domains was obtained by gene cloning techniques,and then recombined into pET-32a(+) vector.The recombinant vector was confirmed by PCR and DNA seqencing analysis.The recombinant plasmid was transformed into E.coli BL21(DE3) and then induced with IPTG for expression.Result] An about 94 kD expected protein was determined by SDS-PAGE.The fusion protein was purified and then immunized New Zealand rabbits,and ITGB6 protein antiserum was prepared.Western blot analysis showed that the antisera could bind to the expressed ITGB6 fusion protein specifically.The ELISA titer of the rabbit anti-ITGB6 sera was 1∶2 560 approximately.Conclusion] Bovine integrin β6 gene is expressed in E.coli.BL21 successfully. |
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| Keywords: | Integrin β6 subunit Prokaryotic expression Antiserum |
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