Animal Husbandry and Veterinary Medicine

Study on Expression and Cell Localization of atrogin-1 Protein in Monopterus albus

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  • (1.College of Biological Science and Engineering,Jiangxi Agricultural University/Jiangxi Agricultural Microbial Resources Development and Utilization Engineering Laboratory,Nanchang 330045,China;2.Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China)

Received date: 2019-11-08

  Online published: 2020-05-15

Abstract

In order to further improve the culture efficiency and muscle quality of Monopterus albus,it is necessary to further explore the genes related to muscle development.In this study,three prokaryotic expression vectors of atrogin-1 were constructed:pCold-TF-atrogin-1,pET28a(+)-atrogin-1 and pGEX6P-1-atrogin-1,and the recombinant vectors were transferred into Transetta(DE3) competent cells. After induced expression of recombinant strains,only pCold-TF-atrogin-1/Transetta(DE3) strain expressed higher soluble recombinant protein.IPTG induction experiment showed that the concentration of inducing agent had little effect on protein solubility and content,and the best induction time was 20 h.The soluble fraction was purified by Ni-NTA column affinity chromatography and SDS-PAGE(10% polyacrylamide) electrophoresis confirmed the recombinant protein.The eukaryotic expression vector pEGFP-N1-atrogin-1 was constructed and the 293T cells were transfected.The experimental results showed that the atrogin-1 fusion protein was expressed in both the cytoplasm and the nucleus,and the expression level was relatively higher in the nucleus.

Cite this article

WANG Yu, WANG Zhaoguang, LIU Zejun, XIA Zhongxi, FAN Yuding, ZHONG Qiwang . Study on Expression and Cell Localization of atrogin-1 Protein in Monopterus albus[J]. Journal of Henan Agricultural Sciences, 2020 , 49(5) : 154 -160 . DOI: 10.15933/j.cnki.1004-3268.2020.05.020

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