基因leg1(liver-enriched gene 1)首先在斑马鱼中作为肝脏富集表达基因被鉴定.进一步的研究揭示leg1编码的Leg1蛋白代表一类新型外分泌蛋白,它在斑马鱼胚期肝脏生长发育过程中起关键作用.小鼠leg1(mu-leg1)是斑马鱼leg1(zb-leg1) 直系向源基因,二者编码的蛋白氨基酸序列相似性为31%.文章通过巢式PCR从成年小鼠肝脏中成功克隆了mu-leg1的cDNA序列,并对该基因在成年小鼠不同组织中的表达特征进行分析和鉴定.Northern印迹杂交和半定量RT-PCR分析结果显示,mu-leg1在成年小鼠小肠中而非肝脏中富集表达.此外,用制备的mu-Leg1多克隆抗体进行Western印迹杂交,结果显示mu-Leg1也是一个分泌蛋白.同时,还建立了mu-leg1基因条件性剔除杂合子小鼠.这些材料为今后深入研究和探讨mu-Leg1蛋白的生化功能奠定了基础.%Gene legl (liver-enriched gene 1) was first identified as a novel gene whose expression was enriched in the liver of zebrafish. Further studies revealed that Legl protein was a novel secretory protein, which played a role in the liver development in zebrafish. Here we reported the analysis of expression pattern of zb-legl homologus gene mu-legl. The cDNA of mu-legl was isolated from adult mouse liver by nested PCR. This gene encodes a putative protein, mu-Legl, which shares 31% similarity with zb-Legl of zebrafish. Both Northern blotting and semi-quantitative RT-PCR demonstrated that the expression of mu-legl was enriched in the small intestine rather than in the liver in adult mouse. We also produced a recombined mu-Legl protein and a mu-Legl specific antibody. Western blottingdemonstrated that mu-Legl was a secretory protein. In addition, we have established a mu-legl conditional knock-out heterozygous mouse. Our work builds a basis for further studies of mu-legl.
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