大约克猪SLA-1原核表达载体的构建及表达

doi:10.16431/j.cnki.1671-7236.2016.12.006

Abstract

Abstract:

In order to construct the prokaryotic expressing vector of SLA-1 derived form Yorkshire pig and express the interest of protein, a pair of primers was designed to amplify the extracellular domain of SLA-1 gene from Yorkshire pig (named SLA-1-DYKe) by PCR. Then the PCR product was cloned into pMD^®19-T Simple Vector and transformed into Escherichia coli TOP10. After cleaved by Nde Ⅰ and Xho Ⅰ, the positive clones were selected to be sequenced. Analyzing by biological soft, the fragment from positive clone with correct sequence was inserted into pET-28a (+) and transformed into E.coli BL21(DE3). After induction and expression, the interest of protein was detected by SDS-PAGE. The results showed that the extracellular domain of SLA-1-DYKe was successfully amplified with the fragment length of 837 bp. The interest of SLA-1 gene was successfully cloned into pMD^®19-T Simple Vector and the positive recombinant plasmids with correct sequences were obtained. The SLA-1-DYKe from positive recombinant plasmids was further inserted into pET-28a(+). After transformed into E.coli BL21(DE3) and induction, the SLA-1-DYKe was successfully expressed. The molecular weight of the protein was about 34 ku. It was concluded that the prokaryotic expressing vector of SLA-1 was constructed successfully from Yorkshire pigs and then the expressed protein was obtained, which would lay a base for studying on the structure and function of SLA-1 from Yorkshire pig in the future.

Key words: Yorkshire pig; SLA-1 gene; prokaryotic expressing vector

CLC Number:

Q786

ZHANG Xiu-juan, YANG Jie, SUN Yong-qiang, GAO Feng-shan. Construction and Expression of SLA-1 Prokaryotic Expressing Vector in Yorkshire Pig[J]. , 2016, 43(12): 3121-3126.

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Construction and Expression of SLA-1 Prokaryotic Expressing Vector in Yorkshire Pig

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