烟台黑猪SLA-2-YTH原核表达载体的构建及表达

Abstract

Abstract: In order to construct the SLA-2-YTH gene prokaryotic expression vector of Yantai Black pig, a pair of primers for amplifying the extracellular domain of SLA-2-YTH by PCR was designed, followed by cloning the gene into pMD 19-T Simple vector, and then the positive clones could be analyzed directly. After cleavage with NdeⅠand XhoⅠ, the positive clone was successfully inserted into pET-28a(+) and then the recombinant plasmids were transformed into competent cell BL21 (Rosseta).After induction, the protein expression could be detected by SDS-PAGE. The results showed that sub-clone of the extracellular domain of SLA-2-YTH was about 834 bp, and it was successfully cloned into pMD 19-T Simple vector showed by the enzyme analysis. By SDS-PAGE, SLA-2-YTH gene was successfully expressed in Escherichia coli BL21 (Rosseta) and the target protein was about 31.0 ku, which was consistent with prediction. After optimization, the relative expressed content of recombinant SLA-2 protein reached more than 25%. Through the research, we successfully constructed the SLA-2-YTH gene prokaryotic expression vector of Yantai Black pig, and then gained the expressed protein, which would lay the foundation for the future study of structure and function of SLA-2.

Key words: Yantai Black pig; swine leukocyte antigen; prokaryotic expression; vector

JIANG Wei, DONG Song-peng, LI Zi-bin, JIANG Long3, FENG Lei, GAO Feng-shan. Construction and Expression of Prokaryotic Expression Vector of SLA-2-YTH Derived from Yantai Black Pig[J]. .

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Construction and Expression of Prokaryotic Expression Vector of SLA-2-YTH Derived from Yantai Black Pig

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