摘要
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 express MH2 domain of smad2 in E.Coli .Methods:The cDNA fragment of MH2 domain was obtained with EcoR I and Xho I from the plasmid pBluescript/S2MH2.The fragment was inserted into prokaryotic gene fusion vector pGEX 4T 2 and an expression plasmid pGEX 4T 2/S2MH2 was constructed. E.Coli DH5 α transformed with recombinant plasmid pGEX 4T 2/S2MH2 was induced by IPTG for 4 h.Results:100 g/L SDS PAGE revealed a new foreign protein band near M r 49×10 3.Conclusion:The constructed plasmid expresses GST MH2 in E.Coli .
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
1999年第2期86-88,共3页
Journal of Practical Stomatology
基金
国家自然科学基金
