Effect of Prokaryotic Expressed Nucleoplasmin on the Efficiency of Somatic Cell Nuclear Transfer in Banna Mini-pig Inbred Line
Effect of Prokaryotic Expressed Nucleoplasmin on the Efficiency of Somatic Cell Nuclear Transfer in Banna Mini-pig Inbred Line
Wenmin Cheng1,2*, Weirong Pan1,2, Yubo Qing1, Yingchao Liu1, Xingqin Zha1,2, Yan Huang2, Jige Xin1,2, Hongjiang Wei1 and Yangzhi Zeng2
ABSTRACT
Nucleoplasmin (Npm) is the most abundant protein in the nucleus of oocytes which can promote in vitro nucleosome formation. In this study, to improve the efficiency of SCNT in Banna Mini-pig Inbred Line (BMI), we obtained the recombinant nucleoplasmin (Npm) by using a prokaryotic expression system and compared the difference on the developmental effects between exogenous Npm and its structural analog, polyglutamic acid (PGA). We chose the pMD18-T vector for Npm expression and transformed E. coli BL21 (DE3) cells to construct a prokaryotic expression system. The results showed that the recombinant Npm protein of 32 kDa could be obtained. Recombinant Npm or PGA was injected into porcine oocytes at different concentrations. The results showed that the blastocyst rates in injection 560 ng/μL Npm and 1000 ng/μL PGA groups were higher than those of other groups. Moreover, the blastocyst rate was highest in injection 560 ng/μL Npm group and was lowest in injection 100 ng/μL PGA group. These data suggested that addition of recombinant Npm obtained by prokaryotic expression or PGA could improve the development of SCNT embryos in the BMI.
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