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大肠杆菌otsA基因的克隆和表达

CLONING AND EXPRESSION OF otsAGENE IN E.COLI

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【作者】 王忆琴戴秀玉王韫恂周坚

【Author】 Wang Yiqin Dai Xiuyu Wang Yunxun Zhou Jian (Institute of microbiology, Chinese Academy of Sciences, Beijing 100080)

【机构】 中国科学院微生物研究所!北京100080

【摘要】 用PCR方法扩增了 1 5kb的otsA基因片段 ,将该片段连接到多拷贝克隆载体后转化otsBA缺失和otsA缺陷的大肠杆菌菌株 ,使转化株重新获得otsA基因功能。生长曲线表明转化株在高渗培养基中生长良好 ,薄层层析法 (TLC)检测海藻糖实验说明转化株细胞经诱导后合成海藻糖 ,otsA基因的克隆和表达为赋予转基因植物抗高渗、耐干旱能力提供了实验依据和材料。

【Abstract】 kb of otsA gene encoding trehalose synthase has been cloned by PCR amplification. The DNA fragment was ligated to multi\|copy vector and transformed to otsBA deleted and otsA deficient strains of \%E.coli\% separately. The transformants exhibited growth as well as the otsBA\++ wild type on medium containing 0.5mol/L NaCl. Trehalose was synthesized and accumulated in the transformed cells under osmotic pressure, which was determined by thin layer chromatograph. The results confirmed that otsA gene was functionally expressed in the recipient strains. These studies suggested that engineering otsA gene and trehalose accumulation into crop plants may improve drought and salinity tolerance.

【关键词】 otsA基因海藻糖PCR扩增表达
【Key words】 otsA geneTrehaloseExpression
【基金】 国家自然科学基金资助项目!( 3 9980 0 2 3 )&&
  • 【文献出处】 微生物学报 ,ACTA MICROBIOLOGICA SINICA , 编辑部邮箱 ,2000年05期
  • 【分类号】Q785
  • 【被引频次】23
  • 【下载频次】139
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