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酿酒酵母海藻糖-6-磷酸合成酶基因克隆及植物表达载体的构建
CLONING OF TREHALOSE -6-PHOSPHATE SYNTHASE GENE FROM S.CEREVISIAE AND ITS PLANT EXPRESSION VECTOR CONSTRUCTION
【摘要】 从耐热性极强的酿酒酵母菌株AS2 1 41 6中分离纯化出总RNA和mRNA ,以AMV逆转录酶合成cDNA ,采用保守引物 ,从该cDNA中扩增克隆出tps1基因 ,对该基因的全序列分析表明 ,该基因含有 1 50 7个核苷酸 ,与国外报道相关基因的同源性达 99 6%。利用BamHⅠ和SacⅠ切点将tps1基因插入植物表达载体 pBin438多克隆位点上 ,得到tps1基因植物表达载体重组质粒。
【Abstract】 Total RNA,mRNA were isolated from baker’s yeast S.cerevisiae , the cDNA was prepared with AMV Reverse Transcriptase from the total mRNA.The gene of trehalose \|6\|phosphate synthase was cloned form the cDNA with PCR amplification. The gene was sequenced and the results showed that the tpsl gene contains 1507 nucleotides and 99.6% identity with S.cerevisiae . The tps1 gene was constructed on the plant expression vector pBin438.
【关键词】 海藻糖-6-磷酸合成酶;
基因克隆;
表达载体;
【Key words】 Trehalose \|6\|phosphate synthase; Gene cloning; Expression vectoD;
【Key words】 Trehalose \|6\|phosphate synthase; Gene cloning; Expression vectoD;
【基金】 国家自然基金资助项目!( 39570 4 0 3)&&
- 【文献出处】 微生物学报 ,Acta Microbiologica Sinica , 编辑部邮箱 ,2001年01期
- 【分类号】Q78
- 【被引频次】11
- 【下载频次】167