中文摘要：

【目的】研究大约克猪SLA-3（SLA-3-YDY）蛋白在体外与口蹄疫病毒多肽的复性。【方法】设计SLA-3-YDY胞外区引物,PCR扩增目的基因,并将此片段克隆至p MD19-T Simple Vector上,双酶切筛选出阳性克隆并测序,测序正确的片段连接至表达载体p ET-21a（＋）上,转化大肠杆菌感受态BL21细胞中,IPTG诱导,SDS-PAGE检测目的蛋白的表达。超声破碎菌体,提取包涵体蛋白,通过稀释复性法将重链SLA-3-YDY、轻链sβ2m和口蹄疫病毒多肽Hu52按摩尔比1：1：1加入复性液,经分子筛层析纯化并检测复合物是否复性。【结果】PCR扩增获得SLA-3-YDY目的基因,经测序阳性克隆序列与原序列一致。经酶切鉴定,证明目的基因与p ET-21a（＋）载体成功连接,经IPTG诱导表达、SDS-PAGE检测,显示目的基因表达,大小约33 k D,SDS-PAGE检测包涵体蛋白,证明包涵体蛋白大小与菌体蛋白大小一致。分子筛及SDS-PAGE检测发现,通过稀释复性法实现了重链、轻链和多肽的复性,并得到蛋白复合物SLA-3-Hu52-sβ2m（45 k D）。【结论】构建了大约克猪SLA-3原核表达载体,获得目的蛋白,实现了口蹄疫病毒多肽Hu52与SLA-3重链及轻链的复性,为今后进一步研究SLA-3的结构和功能奠定基础。

英文摘要：

[Objective] To study the refolding between SLA-3-YDY protein derived from Yorkshire swine and peptides derived from foot-and-mouth disease virus in vitro. [Methods] A pair of primers was designed to amplify the extracellular domain of SLA-3-YDY and then the PCR product was cloned into p MD19-T Simple Vector. After cleaved by Nde I and Xho I, the positive clones were selected to be sequenced. Analyzed by biological soft, the cloned product with correct sequences was selected to be inserted into p ET-21a（＋） and transformed into BL21. After induction with IPTG, the interest of protein was detected by SDS-PAGE. The bacteria were broken ultrasonically, then inclusion body from the bacteria was isolated. The heavy chain of SLA-3-YDY, light chain sβ2m and the epitope-peptides Hu52 from foot-and-mouth disease virus（FMDV）, were refolded at a ratio of 1：1：1 in diluted refolding buffer followed by purification in molecular sieve of superdex 200 to detect whether the complex was refolded. [Results] The PCR result shows that the SLA-3-YDY gene was amplified successfully. After sequencing and analysis, the sequence of the positive clone of SLA-3-YDY was consistent with the primary sequence. By cleavage, the interest of gene was proved to be successfully inserted into p ET-21a（＋） Vector. After induction with IPTG and SDS-PAGE detection, the interest of gene was expressed and the molecular weight was about 33 k D. The isolated inclusion body was also detected by SDS-PAGE, and it was shown that the molecular weight of the inclusion body was consistent with the interest of protein. By refolding in a dilution system, the heavy chains of SLA-3, peptides and light chain succeed to be refolded in vitro. Then, by using the molecular sieve column to separate and purify the refolded proteins and detection by SDS-PAGE, it was shown that the complex protein of SLA-3-Hu52-sβ2m were obtained（45 k D） finally. [Conclusion] The prokaryotic expressing vector of SLA-3 derived from Yorkshire swine was constructed successf