吉富罗非鱼leptin基因的克隆及其表达
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农业部淡水生物多样性保护与利用重点开放实验室第七期开放课题(No.LFBU0706),农业部公益性行业科研专项(No.200903046-2)


Cloning and Expression of leptin in Genetically Improved Farmed Tilapia (GIFT)
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    摘要:

    Leptin为肥胖基因编码产物,是脂肪细胞分泌的蛋白质类激素,在能量平衡、摄食行为调节方面起着重要作用。采用RT-PCR法分离出了吉富罗非鱼(Oreochromis niloticus) leptin基因的部分序列,其cDNA长度为364 bp,编码96个氨基酸。同源性分析显示:鱼类leptin保守性较低。吉富罗非鱼leptin氨基酸序列与斜带石斑鱼(Epinephelus coioides)的相似性较高,为80.6%,但与虹鳟(Oncorhynchus mykiss)、青鳉(Oryzias latipes)、斑马鱼(Danio rerio)等其他鱼的相似性非常低,均在40%以下。使用real time PCR法检测leptin在各种组织中的表达量,结果发现,leptin在肝中相对表达量最为丰富,是肌肉中相对表达量的3 000倍,其次为性腺和脑,分别是肌肉中相对表达量的1 250倍和450倍,而肾、肠和肌肉中表达量较少。

    Abstract:

    Leptin, a product of the obese gene, is a multifunctional hormone secreted predominantly by adipocytes. It plays an important role in regulating energy balance and feeding behavior. Partial cDNA of leptin was isolated from genetically improved farmed tilapia (GIFT) (Oreochromis niloticus) through RT-PCR. The cDNA was 364 bp in length which encoded 96 amino acid residues. Homology analysis of the deduced amino acid sequence of fish leptin was carried out. Fish leptin showed a lower degree of homology. The deduced amino acid sequence of GIFT and Epinephelus coioides leptin was 80.6% identical, however the amino acids homology was below 40% when comparing GIFT with Rainbow Trout (Oncorhynchus mykiss), Medaka (Oryzias latipes), Zebrafish (Danio rerio) and so on. Leptin expression in different tissues was detected by real time PCR. The highest relative expression level was found in liver, which was 3 000 times as high as in muscle; high expression was also found in gonad and brain, which were 1 250 times and 450 times as high as in muscle, respectively. However, the relative expression level was very low in kidney, intestine and muscle.

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唐永凯,俞菊华,徐跑,李建林,李红霞,夏正龙,李创举.2012.吉富罗非鱼leptin基因的克隆及其表达.动物学杂志,47(3):89-94.

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  • 收稿日期:2011-10-28
  • 最后修改日期:2012-03-02
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  • 在线发布日期: 2012-06-22
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