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目的：构建Ｓｍａｄ２基因ＭＨ２结构域原核融合表达载体，在大肠杆菌中表达ＭＨ２结构域，为制备抗ＭＨ２抗体，研究Ｓｍａｄ２基因和ＭＨ２结构域的功能奠定基础。方法：用ｐＧＥＸ－４Ｔ－２表达载体构建ｐＧＥＸ－４Ｔ－２／Ｓ２ＭＨ２原核融合表达载体，转化大肠杆菌ＤＨ５α，ＩＰＴＧ诱导表达ＧＳＴ－ＭＨ２融合蛋白，１００ｇ／ＬＳＤＳ－ＰＡＧＥ检测表达产物。结果：重组载体转化ＤＨ５α，经ＩＰＴＧ诱导４ｈ后，在１００ｇ／ＬＳＤＳ－ＰＡＧＥ上出现一条新的蛋白条带，相对分子质量（Ｍｒ）约为４９×１０３。结论：构建成功ｐＧＥＸ－４Ｔ－２／Ｓ２ＭＨ２融合表达载体，在大肠杆菌ＤＨ５α内表达获得ＧＳＴ－ＭＨ２融合蛋白。
OBJECTIVE: To construct the prokaryotic expression vector of MHC2 domain of Smad2 gene and express MH2 domain in Escherichia coli for the preparation of anti-MH2 antibody and to study the function of Smad2 gene and MH2 domain. Methods: The prokaryotic expression vector pGEX-4T-2 / S2MH2 was constructed by pGEX-4T-2 expression vector and transformed into E. coli DH5α. The expression of GST-MH2 fusion protein was induced by IPTG and the expression product was detected by 100g / LSDS-PAGE. Results: The recombinant vector was transformed into DH5α. After IPTG induction for 4h, a new protein band appeared on 100g / LSDS-PAGE. The relative molecular mass was about 49 × 103. Conclusion: The successful construction of pGEX-4T-2 / S2MH2 fusion expression vector and expression in E. coli DH5α GST-MH2 fusion protein.