摘要
目的 观察釉原蛋白 (amelogenin ,Am)基因聚合酶链反应 (polymerasechainreaction ,PCR)产物在大肠杆菌中的表达。方法 利用谷胱苷肽巯基转移酶表达载体 4T 2型 (ptacglutahionexpression ,PGEX 4T 2 )转化大肠杆菌DH5α ,通过异丙基硫代半乳糖苷 (isopropylthio galactoside ,IPTG)诱导 ,收菌。聚丙烯酰胺凝胶电泳。结果 电泳分析表明 ,釉原蛋白基因PCR产物在大肠杆菌中成功地获得了表达。结论 构建了PGEX 4T 2 /Am融合表达载体 ,获得了融合表达的目的蛋白。
Objective To detect the expression of amelogenin(Am) gene in Escherichia coli (E.coli) . Methods The cDNA fragmain of Am gene was obtained with EcoRI and Xhol I from the plasmid PUC18/Am. The fragment was inserted into prokaryotic gene fusion vector PGEX 4T 2 and an expression plasmid PGEX 4T 2/Am was constructed. PGEX 4T 2/Am was induced by IPTG for 4h. Results 15% SDS PAGE revealed a new forein protein band near 27 000.Conclusion The constructed plasmid expresses Am gene in E.coli.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2001年第1期45-47,共3页
Chinese Journal of Stomatology
