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草鱼leptin基因的分离鉴定及在巴斯德毕赤酵母中的表达
  • 摘要

    利用RT-PCR技术扩增出草鱼(Ctenopharyngodon idellus)的leptin基因,将leptin基因克隆至真核表达载体pPIC9K,电穿孔转化GS115菌株,经G418筛选和甲醇诱导后,对表达产物进行SDS-PAGE琼脂糖凝胶电泳和Western blot分析.结果表明,草鱼leptin基因cDNA序列由438个核苷酸组成,编码146个氨基酸组成的多肽(GenBank登陆号AY551335),与鲤鱼(Cyprinus carpio)leptin基因相比,核苷酸和氨基酸的同源性为99%;与人、猪和鼠相比,核苷酸同源性分别为84%、86%和95%,氯基酸的同源性分别为84%、82%和96%;与河豚(Takifugu rubripes)相比,氨基酸具有较大的差异,仅有9%的同源性,表明leptin在物种的进化上具有一定的差异;实现了草鱼leptin基因在毕赤酵母(Pichia pastoris)中的表达,表达蛋白的分子量约为16 kD,Western blot分析表明,表达产物具有一定的免疫学活性.

  • 作者

    戴汉川  伍晓雄  聂芬  赵娜  张维民  曾翠平  龙良启  DAI Han-chuan  WU Xiao-xiong  NIE Fen  ZHAO Na  ZHANG Wei-min  ZENG Cui-ping  LONG Liang-qi 

  • 作者单位

    华中农业大学动物医学院,武汉,430070/华中农业大学水产学院,武汉,430070

  • 刊期

    2006年5期 ISTIC PKU

  • 关键词

    草鱼  草鱼leptin基因  毕赤酵母表达 

参考文献
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