【摘要】 目的:克隆产麻黄碱重组酵母菌乙醇脱氢酶基因片段,并对其进行序列分析,为研究该基因在重组酵母中与麻黄碱生物合成途径的关系提供参考。方法:根据一段利用抑制差减杂交技术获得的来源于重组酵母乙醇脱氢酶基因片段,采用RACE的方法扩增Adh基因,使用分子生物学软件对该基因进行生物信息学分析。结果:获得一段大小为1 245 bp的基因片段,编码375个氨基酸,含有两个催化域和两个锌结合域,与来源于Candida boidinii ADH3基因的同源性为85%。结论:克隆的基因为乙醇脱氢酶基因,并在GenBank注册,登录号为JF293468。更多还原

【Abstract】 Objective:Clone the recombined yeast alcohol dehydrogenase gene,and analyze the sequence with bioinformatics for the purpose of studying the relation between alcohol dehydrogenase gene and the biosynthesis pathway of ephedrine.Method:Gene special primers were designed based on the fragment of alcohol dehydrogenase from the recombined yeast by suppression subtraction hybridization,the alcohol dehydrogenase gene was obtained by RACE,and molecular biological softwares were used to analyze homologies.Result:The alcohol dehydrogenase gene contains 1 245 bp nucleotide acids,encoding a protein of 375 amino acids,with two catalytic domain and two inserted zinc binding domain.Homology comparison result showed that the alcohol dehydrogenase gene noted a high identy with Candida boidinii ADH3 gene.Conclusion:The cloned fragment was alcohol dehydrogenase gene,and registered into GenBank(accession number: JF293468).更多还原