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| 重组人早孕因子的表达与纯化 | |
| 引用本文： | 杜晓明.重组人早孕因子的表达与纯化[J].中国农学通报,2011,27(14):40-44. |
| 作者姓名： | 杜晓明 |
| 作者单位： | 1. 郑州大学生物工程系,郑州,450001 2. 郑州大学生物工程系,郑州450001;河南省农业科学院,农业部动物免疫学重点开放实验室,河南省动物免疫学重点实验室,郑州450002 3. 河南省农业科学院,农业部动物免疫学重点开放实验室,河南省动物免疫学重点实验室,郑州450002 |
| 摘 要： | 早孕因子(EPF)是具有免疫抑制和生长调节活性的妊娠相关蛋白，为目前最早确认妊娠的生化指标之一。为获得人早孕因子重组蛋白，利用PCR技术扩增人早孕因子基因，克隆入原核表达载体pGEX-6p-1，与GST基因相融合，构建重组表达质粒，转化大肠杆菌表达菌株BL21，经异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达，以SDS-PAGE和Western blot分析重组蛋白的表达，利用GST树脂纯化表达蛋白。结果成功构建重组表达质粒pGEX6P-EPF，在大肠杆菌中诱导表达了GST- EPF融合蛋白，表达蛋白分子量为37.3kDa，并能被抗GST单克隆抗体特异识别，GST亲和层析纯化，制备了EPF重组蛋白。 |
| 关 键 词： | 早孕因子 原核表达 重组蛋白 |
| 收稿时间： | 2011/1/12 0:00:00 |
| 修稿时间： | 2/9/2011 12:00:00 AM |
Prokaryotic Expression and Purification of Early Pregnancy Factor Protein |
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| Du Xiaoming,Zhang Gaiping,Wang Li,Guo Junqing,Wang Linlin,Feng Hua,Quan Kai,Wang Aiping.Prokaryotic Expression and Purification of Early Pregnancy Factor Protein[J].Chinese Agricultural Science Bulletin,2011,27(14):40-44. | |
| Authors: | Du Xiaoming Zhang Gaiping Wang Li Guo Junqing Wang Linlin Feng Hua Quan Kai Wang Aiping |
| Institution: | Du Xiaoming 1,Zhang Gaiping 1,2,Wang Li 2,Guo Junqing 2,Wang Linlin 1,Feng Hua 1,Quan Kai 2,Wang Aiping 1 ( 1 Department of Bioengineering,Zhengzhou University,Zhengzhou 450001,2 Key Laboratory of Animal Immunology of the Ministry of Agriculture,Henan Provincial Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002) |
| Abstract: | Early pregnancy factor (EPF) is a pregnancy-associated protein that has immune-suppressive and growth-regulatory activities. It is regarded as a biochemical index by which super-early pregnancy can be determined. The objective of the study was to express the recombinant EPF protein in Escherichia coli (E. coli). The EPF gene amplified by PCR was sub-cloned into a pGEX-6P-1 vector fused with GST gene to construct a recombinant plasmid. The recombinant plasmid was then transformed into E. coli BL21.The expres... |
| Keywords: | early pregnancy factor (EPF) prokaryotic expression recombinant protein |
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