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| 大肠杆菌O157:H7的sRNA伴侣蛋白Ufq的基因克隆、表达及纯化 | |
| 引用本文： | 刘晓霞,马延,魏波,周围,廖翔,高原,刘虎岐,岳俊杰,梁龙.大肠杆菌O157:H7的sRNA伴侣蛋白Ufq的基因克隆、表达及纯化[J].生物技术通讯,2011,22(2):178-181. |
| 作者姓名： | 刘晓霞 马延 魏波 周围 廖翔 高原 刘虎岐 岳俊杰 梁龙 |
| 作者单位： | 1. 西北农林科技大学,生命科学学院,陕西,杨凌,712100;军事医学科学院,生物工程研究所,北京,100071 2. 军事医学科学院,生物工程研究所,北京,100071 3. 军事医学科学院,生物工程研究所,北京,100071;内蒙古农业大学,动物科学与医学学院,内蒙古,呼和浩特,010018 4. 西北农林科技大学,生命科学学院,陕西,杨凌,712100 |
| 摘 要： | 目的:克隆、表达并纯化肠出血性大肠杆菌(EHEC)O157:H7的sRNA伴侣蛋白Hfq.方法:利用PCR方法从EHEC O157:H7基因组中扩增出基因hfq,并插入含6xHis标签序列的原核表达载体pET28a(+)的多克隆位点中,构建重组表达质粒pET28a(+)-hfq,以重组质粒转化大肠杆菌BL21(DE3)... |
| 关 键 词： | 大肠杆菌O157:H7 sRNA Hfq蛋白 基因克隆 表达 |
Cloning,Expression and Purification of sRNA Chaperone Hfq of EHEC O157:H7 |
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| LIU Xiao-Xia,MA Yan,WEI Bo,ZHOU Wei,LIAO Xiang,GAO Yuan,LIU Hu-Qi,YUE Jun-Jie,LIANG Long.Cloning,Expression and Purification of sRNA Chaperone Hfq of EHEC O157:H7[J].Letters in Biotechnology,2011,22(2):178-181. | |
| Authors: | LIU Xiao-Xia MA Yan WEI Bo ZHOU Wei LIAO Xiang GAO Yuan LIU Hu-Qi YUE Jun-Jie LIANG Long |
| Institution: | LIU Xiao-Xia1,2,MA Yan2,WEI Bo2,3,ZHOU Wei2,LIAO Xiang2,GAO Yuan2,LIU Hu-Qi1,YUE Jun-Jie2,LIANG Long2 1.College of Life Sciences,Northwest A&F University,Yangling 712100,2.Beijing Institute of Biotechnology,Beijing 100071,3.College of Animal Science and Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China |
| Abstract: | Objective：To express and purify small RNA（sRNA） chaperone Hfq of EHEC O157：H7 in Escherichia coli.Methods：The hfq gene coding Hfq protein was amplified by PCR from EHEC O157：H7 genome and inserted into plasmid pET28a（＋） containing 6×His Tag sequence to construct a recombinant vector pET28a（＋）-hfq.The recombinant vector was transformed into host strain E.coli BL21（DE3）.The fusion Hfq was expressed by IPTG induction and detected by SDS-PAGE and Western blot.The fusion protein was purified by Ni2＋-chelating chromatography.Results：The expression vector was digested by restriction enzyme,and the consequent electrophoresis showed the correct length DNA fragment,therefore,the plasmid pET28a（＋）-hfq was correctly constructed.The 17 kD fusion protein expressed in E.coli BL21（DE3） was examined by SDS-PAGE.Finally,the purified fusion protein was successfully obtained from supernatant.Conclusion：The sRNA chaperone Hfq of EHEC O157：H7 was obtained,which will lay the foundation for screening the interacting sRNA and further research on sRNA＇s function. |
| Keywords: | sRNA |
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