| 重组人载脂蛋白AI米兰突变体在大肠杆菌中的高表达 |
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| 引用本文： | 黎明,赵洪亮,薛冲,张伟,张士猛,熊向华,刘志敏. 重组人载脂蛋白AI米兰突变体在大肠杆菌中的高表达[J]. 生物技术通讯, 2003, 14(3): 194-196 |
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| 作者姓名： | 黎明 赵洪亮 薛冲 张伟 张士猛 熊向华 刘志敏 |
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| 作者单位： | 军事医学科学院,生物工程研究所,北京,100071 |
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| 基金项目： | 国家重大科技专项(2002AA2Z345B) |
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| 摘 要： | 用RT-PCR法从人肝总RNA库中克隆出人载脂蛋白Al的cDNA序列，再通过重叠PCR将载脂蛋白AI的第179位精氨酸密码子突变成半胱氨酸密码子，即载胎蛋白AI米兰突变体基因。将此目的基因克隆至表达载体pQE30，重组质粒转化JMl09宿主菌，经表达试验筛选出高表达克隆；工程菌经诱导后表达出含6个氨基酸前肽的载脂蛋白AI米兰突变体。表达产物主要以可溶形式存在，但也有部分为包涵体。
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| 关 键 词： | 载脂蛋白AI米兰突变体 大肠杆菌 表达 重组 |
| 文章编号： | 1009-0002(2003)03-0194-03 |
| 修稿时间： | 2003-01-21 |
High-level expression of recombinant human apolipoprotein A-I-Milano variant in E. coli |
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| LI Ming,ZHAO Hong-liang,XUE Chong,ZHANG Wei,ZHANG Shi-meng,XIONG Xiang-hua,LIU Zhi-min. High-level expression of recombinant human apolipoprotein A-I-Milano variant in E. coli[J]. Letters in Biotechnology, 2003, 14(3): 194-196 |
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| Authors: | LI Ming ZHAO Hong-liang XUE Chong ZHANG Wei ZHANG Shi-meng XIONG Xiang-hua LIU Zhi-min |
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| Abstract: | apolipoprotein A-I-Milano was a natural variant of apolipoprotein A-I.A cDNA of apolipoprotein A-I was amplified from a human liver total RNA library by RT-PCR.Then Arg 173 code in apolipoprotein A-I was mutated into Cys 173 code using PCR technique and this mutated cDNA fragments were subcloned into the expression vector of pQE30.Recombiant plasmids were transformed into E.coli BL21(DE3).The engineered strains of high -level expression of apoliprotein A-I-Milano were selected.The expressed products of apoliprotein A-I-Milano containing a prosegment sequence of six amino acids appeared to be mostly soluble form in E.coli. |
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| Keywords: | apolipoprotein A-I-Milano recombination expression E.coli |
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