| 小麦高分子量谷蛋白亚基1Dx5基因在大肠杆菌中的表达 |
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| 引用本文： | 陈军营,朱雪萍,尹海燕,程西永,许海霞,陈新建.小麦高分子量谷蛋白亚基1Dx5基因在大肠杆菌中的表达[J].麦类作物学报,2007,27(4):597-602. |
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| 作者姓名： | 陈军营 朱雪萍 尹海燕 程西永 许海霞 陈新建 |
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| 作者单位： | 1. 河南农业大学农学院,河南郑州,450002
2. 河南省农业科学院 |
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| 基金项目： | 河南省高校杰出科研创新人才工程项目 |
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| 摘 要： | 为了给小麦优质亚基研究奠定基础,将高分子量谷蛋白亚基1Dx5基因的核苷酸序列与载体pRSET A重组,将构建好的质粒转化大肠杆菌BL21(DE3)pLysS菌株,通过IPTG使其得到了成功表达.大肠杆菌中表达的1Dx5亚基在SDS-PAGE上与小麦品种钱尼中的1Dx5亚基具有相似的迁移率.用Western blot技术成功检测到了目的基因产物.通过改变IPTG浓度、诱导时间、培养基成分及初始菌液浓度研究了最适表达条件.结果表明,最适表达条件为:LB培养基,菌液初始浓度(OD600)0.4～0.6,IPTG浓度0.4mmol/L,诱导时间为3～5 h.
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| 关 键 词： | 小麦 高分子量谷蛋白亚基 大肠杆菌 表达 小麦品种 高分子量 蛋白亚基 基因产物 转化大肠杆菌 表达条件 Gene Wheat Expression 初始浓度 菌液浓度 培养基成分 结果 诱导时间 检测 Western blot 迁移率 相似 IPTG |
| 文章编号： | 1009-1041（2007）04-0597-06 |
| 收稿时间： | 2006-12-11 |
| 修稿时间： | 2006-12-112007-03-05 |
Expression of Wheat HMW-GS 1Dx5 Gene in E. coli |
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| CHEN Jun-ying,ZHU Xue-ping,YIN Hai-yan,CHENG Xi-yong,XU Hai-xi,CHEN Xin-jian.Expression of Wheat HMW-GS 1Dx5 Gene in E. coli[J].Journal of Triticeae Crops,2007,27(4):597-602. |
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| Authors: | CHEN Jun-ying ZHU Xue-ping YIN Hai-yan CHENG Xi-yong XU Hai-xi CHEN Xin-jian |
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| Institution: | College of Agronomy , Henan Agrieutural University,Zhengzhou, Henan 450002,China |
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| Abstract: | Nucleotide acid sequence of HMW-Glu 1Dx5 gene was recombined with the expression vector pRSET A,then the chimaeric gene was transformed into the E.coli strain BL21(DE3) pLysS.The transgenic cells expressed Glu-1Dx5 successively when they were induced by IPTG.1Dx5 subunits expressed in E.coli had the same mobility on SDS-PAGE with the 1Dx5 subunits in dough of wheat cultivar Chineye.The target protein could be visualized successively by analysis of Western blot.Different expression conditions,such as expression times,media,IPTG concentration and cell concentrations(OD600) at the beginning of induction,were tested to optimize culture conditions.In LB medium,good expression effects were obtained at culture conditions of 0.4 mmol/L IPTG,cell concentration of 0.4~0.6(OD600) and 3~5 h induction time. |
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| Keywords: | High molecular weight glutenin subunits E coli Expression |
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