摘要
目的：观察釉原蛋白（Ａｍｅｌｏｇｅｎｉｎ，Ａｍ） 基因ＰＣＲ 产物在大肠杆菌中的表达。方法：利用原核表达载体ＰＲＳＥＴ，转化大肠杆菌ＪＭ１０９ ，通过ＩＰＴＧ 诱导，收菌。聚丙烯酰胺凝胶电泳。过柱纯化。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的获得了表达。结论：表达的融合蛋白主要以包涵体的形式存在。利用Ｎｉ－ ＮＴＡ 金属螯合亲和层析在变性条件下对表达的蛋白进行纯化，获得纯化蛋白。
Aim: To express Am gene in E.coli . Methods: The cDNA fragment of Am gene was obtained with EcoR Ⅰ and Hind Ⅲ from the plasmid PUC18/Am.The fragment was inserted into prokaryotic gene fusion vector PRSET and an expression plasmid PRSET/Am was constructed.PRSET/Am was induced by IPTG for 4h. Then metal chelate interaction chromatography was used to purify protein. Results: 150g/L SDS-PAGE revealed a new foreing protein band near Mr 24000. Conclusions: The constructed plasmid expression Am gene in E.coli .
出处
《牙体牙髓牙周病学杂志》
CAS
1999年第4期261-263,共3页
Chinese Journal of Conservative Dentistry
关键词
釉原蛋白
基因表达
融合
纯化
Ni-NTA
制备
Amelogenin, expression, purification, metal chelate interaction chromatography
