| 蜡梅金属硫蛋白基因的克隆与原核表达 |
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| 引用本文： | 秦平伟,段明革,孙文婷,李名扬,徐晶.蜡梅金属硫蛋白基因的克隆与原核表达[J].北京林业大学学报,2010(Z2). |
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| 作者姓名： | 秦平伟 段明革 孙文婷 李名扬 徐晶 |
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| 作者单位： | 西南大学园艺园林学院;重庆市花卉工程技术研究中心;重庆城市管理职业学院; |
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| 基金项目： | 重庆市教委科技项目(KJ070215) |
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| 摘 要： | 金属硫蛋白(metallothionein,MT)是一类富含Cys,能够结合重金属的低分子量蛋白质,广泛分布于生物界。在构建好的蜡梅花cDNA文库并进行EST分析的基础上,通过随机克隆测序得到了1个蜡梅金属硫蛋白的cDNA,命名为CpMetallothionein(CpMT)。CpMTcDNA全长为1083bp,基因内部含有一长度为240bp的开放阅读框,可编码79个氨基酸残基。将CpMT插入原核表达载体pET-32a,并转化Origami2感受态细胞。诱导表达产物经SDS-PAGE结果显示,目的蛋白约为30kD。表达的融合蛋白以包涵体和可溶性蛋白2种形式存在,用His-Bind蛋白纯化回收试剂盒对其进行纯化回收,得到了高纯度蛋白,为今后研究奠定基础。
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| 关 键 词： | 金属硫蛋白 克隆 序列分析 原核表达 |
Cloning and prokaryotic expression of a metallothionein cDNA from Chimonanthus praecox (L.) Link |
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| QIN Ping-wei,DUAN Ming-ge,SUN Wen-ting,LI Ming-yan g,XU Jing. College of Horticulture , L,scape Architecture,Southwest University,Chon gqing,P.R.China, Chongqing Engineering Research Center for Floriculture, Chongqing City Management College,P.R.China..Cloning and prokaryotic expression of a metallothionein cDNA from Chimonanthus praecox (L.) Link[J].Journal of Beijing Forestry University,2010(Z2). |
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| Authors: | QIN Ping-wei DUAN Ming-ge SUN Wen-ting LI Ming-yan g XU Jing College of Horticulture L scape Architecture Southwest University Chon gqing PRChina Chongqing Engineering Research Center for Floriculture Chongqing City Management College PRChina |
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| Institution: | QIN Ping-wei1,2,DUAN Ming-ge3,SUN Wen-ting1,LI Ming-yan g1,XU Jing3.1 College of Horticulture and Landscape Architecture,Southwest University,Chon gqing,400715,P.R.China,2 Chongqing Engineering Research Center for Floriculture,3 Chongqing City Management College,400055,P.R.China. |
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| Abstract: | Metallothioneins(MT) are low-molecular-weight proteins combined with heavy metals,which are rich in Cys and widely distributed in the biosphere.In this paper,a MT gene,designated as CpMetallothionein(CpMT),was obtained by sequencing the randomly selected clones,on the basis of cDNA library construction of Chimonanthus praecox(L.) Link and ESTs analysis.The bioinformation analysis showed that the cDNA sequence has a length of 1 083 bp,containing ORF of 240 bp which encodes 79 amino acids.To get the fusion pr... |
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| Keywords: | metallothionein cloning sequence analysis prokaryotic expression |
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