摘要
利用酵母密码子偏爱性将黑曲霉(Aspergillus niger)中的内切菊粉酶(Endoinu linase)基因通过基因全合成的方式合成为酵母密码子偏爱性的内切菊粉酶基因。然后将原始和全合成的内切菊粉酶基因克隆到解脂耶氏酵母表达载体PINA1296上,得重组解脂耶氏酵母表达载体pHBM2020、pHBM2021,将两种质粒分别转化解脂耶氏酵母(Yarrowia lipolytica)CLIB725,筛选得到重组解脂耶氏酵母CLIB725(pHBM2020)、CLIB725(pHBM2021),将两种重组酵母摇瓶培养,经SDS-PAGE、测酶活检测表明两种基因在解脂耶氏酵母中都有表达,全合成菊粉酶比原始菊粉酶酶活要高。
By using the principle of preference of the yeast codon, the endoinulinase gene was changed into its synthesis gene of preference of the yeast codon. These two genes were cloned into the vector of the expression of Yarrowia lipolytica pINA1296, and the recombinant vectors pHBM2020 ,pHBM2021 were obtained. The two recombinant vectors were transferred into Yarrowia lipolytica CLIB725, and the two recombinant strains CLIB725 （pHBM2020） ,CLIB725 （pHBM2021）were constructed. The two strains were cultured in the flask. And the analysis of the SDS -PAGE and the enzyme activity showed that the two genes were expressed in Yarrowia lipolytica, and the enzyme activity of synthesis gene of the endoinulinase was higher than that of the original endoinulinase gene.
出处
《氨基酸和生物资源》
CAS
2006年第2期32-34,44,共4页
Amino Acids & Biotic Resources
