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| Constructing Mouse pET3C-Myc Vector and Its Expression in Rosetta(DE3) and Its Purification |
| XIANG Li1, WANG Shen2, TIAN Hai-shan2, ZHONG Mei-juan1, ZHOU Ru-bin1, CAO Ding-guo1, LIANG Peng1, ZHANG Guo-ping1, HE Tao1, PANG Shi-feng1 |
1. Department of Biology, Guangdong Medical University, Dongguang 523808, China;
2. School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325035, China |
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Abstract Objective: The object is to construct prokaryotic expression vector which carried mouse cloned c-Myc gene, and achieve its expression and then purify the product.Methods: TetO-FUW-OSKM plasmid was used as a templet to amplify c-Myc gene fragment by polymerase chain polymerization reaction (PCR). After inserting into pET3C vector, the c-Myc gene was transformed into E.coli DH5α. The eligible recombinant plasmid pET3C-Myc was screened out after resistance selection, restrictive identification by PCR, and sequencing and then transformed into E.coli Rosetta (DE3).Result: The result of DNA agarose gel electrophoresis show the amplified gene met the size of destination gene. The sequencing of the reconstructed gene was the same as that of in GenBank. Reaching the destination, the purified product proved to have about 64kDa by SDS-PAGE test. Conclusion: It is successful to construct the recombinant plasmid pET3C-Myc, and achieve its expression in the Rosetta (DE3) and purify the product. The above success paves the path for next study.
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Received: 04 November 2016
Published: 25 February 2017
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Cite this article:
XIANG Li, WANG Shen, TIAN Hai-shan, ZHONG Mei-juan, ZHOU Ru-bin, CAO Ding-guo, LIANG Peng, ZHANG Guo-ping, HE Tao, PANG Shi-feng. Constructing Mouse pET3C-Myc Vector and Its Expression in Rosetta(DE3) and Its Purification. China Biotechnology, 2017, 37(2): 20-25.
URL:
https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20170204 OR https://manu60.magtech.com.cn/biotech/Y2017/V37/I2/20
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