人釉原蛋白编码区基因真核表达载体PsecTaq2A-AMG的构建

华西口腔医学杂志

人釉原蛋白编码区基因真核表达载体PsecTaq2A-AMG的构建

杨爱玲,徐琛蓉,章锦才,钟良军,殷春一,赵川江

610041　四川大学华西口腔医学院口腔内科学教研室 (杨爱玲,徐琛蓉,殷春一,赵川江),广东省口腔医院口腔内科(章锦才),新疆医科大学第一附属医院口腔科(钟良军)

收稿日期:2003-04-25 修回日期:2003-04-25 出版日期:2003-04-20 发布日期:2003-04-20

Construction of the Eukaryotic Expression Vector PsecTaq2A-AMG for Human Amelogenin

YANGAiling*,XUChenrong,ZHANGJincai,et al.

*OralMedicine ofWest China College ofStomatology,Sichuan Universi- ty,Chengdu610041,China

Received:2003-04-25 Revised:2003-04-25 Online:2003-04-20 Published:2003-04-20

摘要/Abstract

摘要：

目的　构建人釉原蛋白(AMG)编码区基因真核表达载体PsecTaq2A-AMG。方法　采用PCR技术体外扩增AMG完整分泌肽编码区。将扩增产物与PsecTaq2A分别用BamHI和Xhol I行双酶切,将获取的AMG目的基因片段连接到双酶切后的PsecTaq2A,构建重组质粒PsecTaq2A-AMG,并对重组质粒进行鉴定。结果　①PCR扩增产物经1·5%琼脂糖凝胶电泳,可见大小约519 bp的特异性条带,与预期结果一致。②重组克隆PsecTaq2A-AMG酶谱分析与预期结果一致,序列测定结果与GenBank中的人釉原蛋白序列完全一致。结论　用此方法可成功构建AMG 编码区基因真核表达载体PsecTaq2A-AMG。

关键词: 人釉原蛋白, 重组质粒, PsecTaq2A-AMG

Abstract:

Objective　The purpose of this study was to construct a eukaryotic expression vector for human amelogenin (AMG).Methods　PCR was performed to amplify the AMG encoding region. Amplified fragments for human AMG were recov- ered and inserted into eukaryotic expression vectors PsecTaq2A. The recombinant plasmid PsecTaq2A-AMG was constructed and their positive cloneswere identified.Results　①Amplified productswere checked by electrophoresis and the resultswere satisfac- tory.②The recombinant plasmid PsecTaq2A-AMG was analyzed by restriction endonuclease mapping and DNA sequencing. The results of sequencing were consistentwith those fromGenBank.Conclusion　The recombinant plasmid PsecTaq2A-AMGwas suc- cessfully constructed with properly inserted DNA sequence encoding mature amelogenin.

Key words: human amelogenin, recombinant plasmid, PsecTaq2A-AMG

杨爱玲,徐琛蓉,章锦才,钟良军,殷春一,赵川江. 人釉原蛋白编码区基因真核表达载体PsecTaq2A-AMG的构建[J]. 华西口腔医学杂志.

YANGAiling*,XUChenrong,ZHANGJincai,et al.. Construction of the Eukaryotic Expression Vector PsecTaq2A-AMG for Human Amelogenin[J]. West China Journal of Stomatology.

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