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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.

Key words: Monopterus albus, atrogin-1, Prokaryotic expression, Purification, Cell transfection, Cell localization