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盐藻磷酸甘油脱氢酶基因cDNA文库的构建

cDNA CLONING OF GLYCEROL3PHOSPHATE DEHYDROGENASE GENE OF DUNALIELLA SALINA

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【作者】 周冀明白林含吴晓蓉蒋彦王峰刘世贵

【Author】 Zhou Jiming Bai Linhan Wu Xiaorong Jiang YanWang Feng Liu Shigin(National Laboratory of Grassland Biotechnology))

【机构】 草原生物工程国家专业实验室

【摘要】 采用Lambdagt10载体构建了盐藻cDNA文库,文库大小为106/mL,插入片段平均长度大于1kb.根据果蝇、兔、鼠编码其3-磷酸甘油脱氢酶基因的辅酶NAD的结合功能区保守序列设计一对引物,大小分别为21bp和18bp.PCR扩增得到与果蝇相同的290bp特异扩增片段,以该片段为探针,采用缺口平移系统标记,原位杂交,从盐藻cDNA文库中获得36个3-磷酸甘油脱氢酶基因的阳性克隆

【Abstract】 The cDNA library (106 clones) of Dunaliella Salina was obtained using a lambda gt10 vector.The average size of insertion is more than 1kb.Comparing the conservative sequence encoding NAD binding region of glycerol3phosphate dehydrogenase among Drosophila,rabbit and mouse,we designed and synthesized a pair of primers.The size of the primers were 21bp and 18bp respectively.PCR amplification was performed and obtained ene specific amplified fragment of 290bp which is as same as in Drosophila DNA.The fragment was recoveried and used as the probe for screening glycerol3phosphate dehyogenase gene from cDNA library.Nick translation system was used for labelling probe.As a result of in situ hybrdization,36 positive clones were obtained.

【基金】 国家自然科学基金
  • 【文献出处】 四川大学学报(自然科学版) ,Journal of Sichuan University (Natural Science Edition) , 编辑部邮箱 ,1997年06期
  • 【分类号】Q781
  • 【被引频次】26
  • 【下载频次】85
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