斑马鱼leg1直系同源基因mu-leg1在小鼠中的表达谱分析

doi:10.3724/SP.J.1005.2012.01174

遗传 ›› 2012, Vol. 34 ›› Issue (9): 1174-1180.doi: 10.3724/SP.J.1005.2012.01174

斑马鱼leg1直系同源基因mu-leg1在小鼠中的表达谱分析

朱智慧, 胡敏杰, 常长青, 彭金荣

浙江大学动物科学学院, 杭州 310058

收稿日期:2012-03-13 修回日期:2012-04-06 出版日期:2012-09-20 发布日期:2012-09-25
通讯作者: 彭金荣 E-mail:pengjr@zju.edu.cn
基金资助:
国家杰出青年科学基金项目(编号：30825025)资助

Analysis of expression pattern of zebrafish leg1 homologus gene mu-leg1 in mouse

ZHU Zhi-Hui, HU Min-Jie, CHANG Chang-Qing, PENG Jin-Rong

College of Animal Sciences, ZhejiangUniversity, Hangzhou 310058, China

Received:2012-03-13 Revised:2012-04-06 Online:2012-09-20 Published:2012-09-25

摘要/Abstract

摘要： 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)的直系同源基因, 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。

关键词: mu-leg1, 重组蛋白, 多克隆抗体, 条件性基因剔除小鼠

Abstract: Gene leg1 (liver-enriched gene 1) was first identified as a novel gene whose expression was enriched in the liver of zebrafish. Further studies revealed that Leg1 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-leg1 homologus gene mu-leg1. The cDNA of mu-leg1 was isolated from adult mouse liver by nested PCR. This gene encodes a putative protein, mu-Leg1, which shares 31% similarity with zb-Leg1 of zebrafish. Both Northern blotting and semi-quantitative RT-PCR demonstrated that the expression of mu-leg1 was enriched in the small intestine rather than in the liver in adult mouse. We also produced a recombined mu-Leg1 protein and a mu-Leg1 specific antibody. West-ern blottingdemonstrated that mu-Leg1 was a secretory protein. In addition, we have established a mu-leg1 condi-tional knock-out heterozygous mouse. Our work builds a basis for further studies of mu-leg1.

Key words: mu-leg1, recombinant protein, polyclonal antibody, conditional knock-out mouse

朱智慧，胡敏杰，常长青，彭金荣. 斑马鱼leg1直系同源基因mu-leg1在小鼠中的表达谱分析[J]. 遗传, 2012, 34(9): 1174-1180.

ZHU Zhi-Hui, HU Min-Jie, CHANG Chang-Qing, PENG Jin-Rong. Analysis of expression pattern of zebrafish leg1 homologus gene mu-leg1 in mouse[J]. HEREDITAS, 2012, 34(9): 1174-1180.

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斑马鱼leg1直系同源基因mu-leg1在小鼠中的表达谱分析

Analysis of expression pattern of zebrafish leg1 homologus gene mu-leg1 in mouse

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