人釉原蛋白编码区基因真核表达载体PsecTaq2A-AMG的构建
Construction of the Eukaryotic Expression Vector PsecTaq2A-AMG for Human Amelogenin
作者：杨爱玲,徐琛蓉,章锦才,钟良军,殷春一,赵川江
Author：YANGAiling*,XUChenrong,ZHANGJincai,et al.
收稿日期：2003-04-25 年卷（期）页码：2003,21(02):133-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：人釉原蛋白,重组质粒,PsecTaq2A-AMG,
Key words：human amelogenin,recombinant plasmid,PsecTaq2A-AMG,
基金项目：
中文摘要
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。
英文摘要
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.
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