| 富含蛋氨酸玉米醇溶蛋白在大肠杆菌中的表达 |
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| 引用本文： | 娄丽,高学军.富含蛋氨酸玉米醇溶蛋白在大肠杆菌中的表达[J].生物技术,2011,21(5). |
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| 作者姓名： | 娄丽 高学军 |
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| 作者单位： | 东北农业大学乳品科学教育部重点实验室,黑龙江哈尔滨,150030 |
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| 基金项目： | 黑龙江省农业生物功能基因重点实验室开放课题 |
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| 摘 要： | 目的:克隆玉米中富含蛋氨酸的10kD玉米醇溶蛋白,证明植物源目的基因在大肠杆菌中能够表达.方法:从玉米胚乳中克隆高蛋氨酸基因zein,通过PCR扩增zein片段,连接pGEX - 4T -1原核表达载体,转入大肠杆菌中,IPTG诱导后,HPLC测定蛋氨酸含量.结果:经PCR扩增出467bp条带,Blast分析同源性99%,经IPTG诱导进行SDS - PAGE检测,发现在36kD处出现一条明显的条带.诱导后菌体总蛋氨酸含量比正常菌体提高了9.6%.结论:证实了植物源10kD玉米醇溶蛋白在大肠杆菌中能够表达.
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| 关 键 词： | 高蛋氨酸蛋白基因 玉米醇溶蛋白 大肠杆菌 表达 |
Expression of Methionine -rich Zein in E.coli |
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| LOU Li,GAO Xue-jun.Expression of Methionine -rich Zein in E.coli[J].Biotechnology,2011,21(5). |
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| Authors: | LOU Li GAO Xue-jun |
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| Institution: | LOU Li,GAO Xue-jun*(Key Laboratory of Dairy Science of Ministry of Education,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | Objective:To make sure the expression of interest gene rich-methionine 10kD zein in E.coli.Method:The methionine-rich protein gene zein was cloned from maize endosperm.Zein gene obtained by PCR was cloned,constructed with prokaryotic expression vector pGEX-4T-1,and then transferred to E.coli,tested by SDS-PAGE after IPTG inducing.The methionine content of recombination bacterium was tested by HPLC.Result: The 467bp band was amplied by PCR and its homology was 99% by blast.Upon IPTG induction,an apparent app... |
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| Keywords: | methionine-rich gene zein E coli expression |
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