Transforming growth factor-β1 type Ⅱreceptor(TGFⅡR)is the main receptor of TGF-β family.Existing studies have shown that TGF-β1 plays a negative regulatory role in livestock reproduction activities.There-fore,we proposed a new idea to improve the reproductive performance of livestock by competitively binding TGF-β1 in vivo with recombinant TGFⅡR extracellular domain protein.In order to verify it,this study first optimized the codon of porcine TGFⅡ R extracellular domain and syn-thesized it artificially.The synthesized gene fragment was inserted into the prokaryotic expression vector pET-32a(+).Then,the recombinant expression vector was transferred into BL21(DE3)expression strain,and suitable lactose induction concentration and induction time were screened to obtain the highest expression efficiency.The recombinant protein was pu-rified,regenerated and identified by mass spectrometry.Finally,the bioactivity of the recombinant protein was determined by in vitro cell experiment.The results showed that the high expression of recombinant protein was induced at 37℃,6 h,2.0 g/L lactose,the inclusion body protein was dissolved by 8 mol/L urea,and the recombinant porcine TGFⅡR protein with a purity of more than 80%was obtained after dialysis with normal saline for 24 h.After treatment of porcine granulosa cells with the recombinant protein,the phosphorylation level of Smad3 induced by TGF-β1 was significantly inhibited,which indicated that the recombinant porcine TGFⅡR protein expressed in this study had good biological activity.In con-clusion,this study lays a foundation for further research and development of products suitable for improving the reproductive efficiency of pigs.
维普
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