Construction and Expression of Prokaryotic Expression Vector of SLA-2-YTH Derived from Yantai Black Pig |
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| Authors: | JIANG Wei DONG Song-peng LI Zi-bin JIANG Long FENG Lei GAO Feng-shan |
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| Institution: | (1. College of Life Science and Technology, Dalian University, Dalian 116622, China; 2. College of Life Science, Jilin Agricultural University, Changchun 130118, China; 3. Zhuolu County Hospital in Hebei Province, Zhuolu 075600, China） |
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| 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. |
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| Keywords: | Yantai Black pig swine leukocyte antigen prokaryotic expression vector |
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