以托佩克猪重链基因SLA-2和轻链基因β2m为研究对象,体外通过剪接重叠延伸PCR(spli-cing overlap extension PCR,SOE-PCR)技术重构复合体,并将复合体基因链克隆入p2X表达质粒,导入受体菌TB1并进行表达。结果显示,体外重构的托佩克猪SLA-2与β2m复合体基因以融合蛋白MBP-SLA-2-(G4S)3-β2m的形式表达,大小为84.1ku。为进一步在体外进行该品系猪复合体与多肽结合研究奠定了基础。
Aimed as the heavy chain gene SLA-2 and the light chain gene β2m from Topics pigs,the complex was reconstructed with splicing overlap extension PCR(SOE,PCR) followed by cloning the complex gene into the expressing vector p2X and transforming the recombinant vector into E.coli.TB1 to be induced to express.The result indicated that the reconstructed complex gene was successfully expressed as a fusion protein MBP-SLA-2-(G4S)3-β2m with 84.1 ku in vitro.This research will pave the way to use the TOPICS swine SLA-2 and β2m complex to bind peptides in vitro.