pVT102U/α-bgln重组真核表达质粒的构建和鉴定  

Construction and Identification of the Recombinant Eukaryotic Expression Plasmid pVT102U/α-bgln

作　　者：林志楷[1] 叶冰莹[1] 潘海芳[1] 陈由强[1] 陈如凯[2] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108 [2]农业部甘蔗生理生态与遗传改良重点开放实验室,福建福州350108

出　　处：《福建师范大学学报（自然科学版）》2007年第2期71-74,共4页Journal of Fujian Normal University：Natural Science Edition

基　　金：国家自然科学基金资助项目(30371177);福建省科技厅重点项目(2005N037);卫生部基金资助项目(JA05225/wkj2);福建省自然科学基金资助项目(B0310004)

摘　　要：构建重组真核表达质粒pVT102U/α-bgln使之可以在酿酒酵母中表达以获得β-葡糖苷酶,用于提高从植物中提取白藜芦醇的得率.以含有从黑曲霉中获得的编码β-葡萄糖苷酶(bgln)成熟肽的cDNA的克隆质粒pMD19T-bgln为模板,用PCR的方法扩增β-葡糖苷酶基因片段(bgln),利用真核表达载体pVT102U/α构建pVT102U/α-bgln重组质粒,PCR结果显示扩增片段约2.6 kb,与预期相同.用酶切电泳验证重组结果的正确性,重组质粒酶切后显示其大小约10 kb,大小及酶切图谱与预期相同.测序检测质粒重组后序列情况,经测序发现插入片段两端序列无改变,且读码框正确.pVT102U/α-bgln质粒构建成功.A recombinant plasmid pVT102U/α-bgln was constructed for obtaining the BGLN protein expressed in Saccharomyces Cerevisiae so as to develop the efficiency of extracting resveratrol from plants.PCR was used to amplify the bgln gene from plasmid pMD19T-bgln in which bgln was cloned from Aspergillus Niger.A new plasmid pVT102U/α-bgln,was then constructed by inserting the amplified bgln gene into pVT102U/α,a eukaryotic expression vector.Restriction analysis and sequencing were used to confirm the structure of pV...

关 键 词：Β-葡萄糖苷酶 pVT102U/α 构建 真核表达 

分 类 号：Q78[生物学—分子生物学]