| 盐生植物盐穗木HcPS_H基因大肠杆菌表达载体的构建 |
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| 引用本文： | 郝晓燕,张毓露,足木热木,刘小利.盐生植物盐穗木HcPS_H基因大肠杆菌表达载体的构建[J].安徽农业科学,2012,40(10):5758-5759. |
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| 作者姓名： | 郝晓燕 张毓露 足木热木 刘小利 |
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| 作者单位： | 新疆农业科学院核技术生物技术研究所,新疆乌鲁木齐,830091;新疆农业大学农学院,新疆乌鲁木齐,830052 |
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| 基金项目： | 新疆维吾尔自治区农作物生物技术重点实验室开放课题(XJDX0201-2011-05) |
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| 摘 要： | 目的]构建盐穗木HcPS_H基因的大肠杆菌表达载体。方法]以质粒pGEM-T-HcPS_H为模板,HcPS_H-pET28a-F、HcPS_H-pET28a-R为引物进行PCR扩增,获得两端含酶切位点的HcPS_H基因目的片段。将该目的片段与大肠杆菌表达载体pET-28a通过T4DNA连接酶连接,并转化大肠杆菌(Escherichia coli)DH5α,得到重组子克隆,抽质粒进行测序鉴定。结果]扩增出HcPS_H基因片段长度为441 bp,将其连接到大肠杆菌表达载体pET-28a上后,得到重组子质粒,经PCR检测、双酶切验证及测序鉴定,表明成功构建原核表达载体。结论]该研究成功构建了盐穗木HcPS_H基因的大肠杆菌表达载体,为进一步进行该基因功能的研究奠定了基础。
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| 关 键 词： | 盐生植物 盐穗木 HcPS_H基因 原核表达载体 |
Constructing of a E.coli Expression Vector of HcPS_H Gene from Halostachys caspica |
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| Institution: | HAO Xiao-yan et al(Institute of Nuclear and Biological Technologies,Xinjiang Academy of Agricultural Sciences,Urumqi,Xinjiang 830091) |
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| Abstract: | Objective]To construct a E.coli expression vector of HcPS_H gene from Halostachys caspica.Method]The HcPS_H gene full-length fragment was obtained from pGEM-T-HcPS_H by PCR using a pair of including enzyme site primer,and then the purified fragment was cloned into pET-28a vector.Result]HcPS_H gene was amplified from Halostachys caspica.The gene fragment was 441bp,and was cloned into pET-28a vector.After analysis of restriction endonuclease digestion and PCR,the constructed expression vector was proved to be correct.Conclusion]The expression vector of HcPS_H gene of Halostachys caspica was constructed successfully. |
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| Keywords: | Halophytes Halostachys caspica HcPS-H gene E coli expression vector |
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