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| Molecular Cloning, Prokaryotic Expression and Polyclonal Antibody Preparation of Peroxiredoxin 6 (Prdx6) from Mus musculus |
| CHEN Xiao-feng1, HU Pan1, LI Yan-song1, GUO Xing2, ZOU De-ying2, LIU Nan-nan1, LU Shi-ying1, ZHOU Yu1, LIU Zeng-shan1, LI Zhao-hui1, REN Hong-lin1 |
1. Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China;
2. Panjin Inspection and Testing Center, Panjin 124010, China |
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Abstract Peroxiredoxin6 (Prdx6) is a bifunctional protein that includes glutathione peroxidase and calcium-independent phospholipase A2. A SSH cDNA library of buffy coat from Ovis aries was constructed through infecting with virulent and avirulent strains of Brucella, and a differentially expressed Prdx6 was cloned.It indicated that Prdx6 might take part in the infection of brucella. In order to study the Prdx6 protein function, here total RNAs from the mice macrophage cell line Raw 264.7 were extracted, cDNAs were reverse transcribed, and the open read frame (ORF) of mouse Prdx6 gene was cloned using the specific primes by means of RT-PCR. Then, the prokaryotic expression vector of Prdx6 was constructed, and the recombinant Prdx6 protein with a His tag was expressed by induction of IPTG, and purified by affinity chromatography of Ni-NTA. A specific polyclonal antibody against mouse Prdx6 was obtained by immunizing a rabbit with the purified Prdx6, which would benefit the further study on the functions of Prdx6 protein in the future.
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Received: 23 September 2015
Published: 01 December 2015
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Cite this article:
CHEN Xiao-feng, HU Pan, LI Yan-song, GUO Xing, ZOU De-ying, LIU Nan-nan, LU Shi-ying, ZHOU Yu, LIU Zeng-shan, LI Zhao-hui, REN Hong-lin. Molecular Cloning, Prokaryotic Expression and Polyclonal Antibody Preparation of Peroxiredoxin 6 (Prdx6) from Mus musculus. China Biotechnology, 2016, 36(3): 11-16.
URL:
https://manu60.magtech.com.cn/biotech/DOI:10.13523/j.cb.20160302 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I3/11
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