短小芽孢杆菌C-9葡聚糖内切酶基因的克隆及其在酿酒酵母中的表达

从短小芽孢杆菌(Bacillus pumilus)C-9中克隆得到葡聚糖内切酶基因,该基因包含1980 bp核苷酸,编码559个氨基酸,其N端有一个预测的29个氨基酸的信号肽序列;以YIp5质粒为骨架,构建rDNA介导的多拷贝整合载体,实现葡聚糖内切酶在酿酒酵母中的高效、稳定表达。与对照菌株相比,含有葡聚糖内切酶基因的重组酿酒酵母可以在以羧甲基纤维素为唯一碳源的培养基上生长。

Cloning of glucanase gene from Bacillus pumilus C-9 and its expression in Saccharomyces cerevisiae

The glucanase gene was obtained from Bacillus pumilus C-9,the open reading frame of glucanase consists of 1 980 nucleotides encoding a protein of 559 amino acids with a predicted signal peptide of 29 amino acids at the N-terminal.To achieve the stable and high efficient expression in Saccharomyces cerevisiae,a multicopy integration plasmid was constructed by introducing the rDNA sequence into the plasmid YIp5.The recombinant Saccharomyces cerevisiae could grow on the plates with carboxymethyl cellulose as the sole carbon source.