猪Prox1基因的染色体定位和亚细胞定位的研究

畜牧兽医学报

猪Prox1基因的染色体定位和亚细胞定位的研究

袁继红，龙欢，田明芳，杨在清^*

(华中农业大学生命科学技术学院，武汉 430070)

收稿日期:2012-07-16 出版日期:2013-03-23 发布日期:2013-03-23
通讯作者: 杨在清，教授，博导，主要从事动物脂肪沉积分子机理研究，E-mail：yangzq@mail.hzau.edu.cn
作者简介:袁继红（1967-），女，湖北武汉人，硕士，实验师，主要从事动物脂肪沉积分子机理研究， E-mail：jhjhyz@mail.hzau.edu.cn
基金资助:
国家自然科学基金项目“SIRT1对猪骨髓间充质干细胞成脂分化的调控机制”（30771585）

Study on Chromosomal and Subcellular Localization for Porcine Prox1 Gene

YUAN Ji-hong, LONG Huan, TIAN Ming-fang, YANG Zai-qing^*

（College of Life Science and Technology，Huazhong Agricultural University，Wuhan 430070, China）

Received:2012-07-16 Online:2013-03-23 Published:2013-03-23

摘要/Abstract

摘要：

摘要：在克隆猪Prox1基因（sProx1）的基础上，对猪Prox1基因进行染色体定位分析，并通过EGFP融合蛋白对该基因产物进行亚细胞水平定位。本研究将人和鼠的Prox1基因编码区在猪的ESTs数据库中进行检索，然后根据EST进行序列拼接并设计引物对猪Prox1基因编码区进行克隆。通过比对人、鼠及其他物种Prox1基因第二个内含子序列，设计引物扩增猪Prox1基因部分内含子，根据克隆得到的部分内含子序列设计引物，利用猪仓鼠体细胞辐射杂种板（IMpRH）对猪Prox1基因进行染色体定位。根据已获得的猪Prox1基因编码区序列，构建猪Prox1基因的融合绿色荧光超表达载体EGFP-Prox1，并转染猪肾PK15细胞，12 h后倒置荧光显微镜观察Prox1蛋白在亚细胞内的定位。结果表明：Prox1 基因定位于猪第9号染色体长臂的26区，与标记SW1651紧密连锁；构建猪Prox1基因的真核表达载体pEGFP -N1- Prox1，融合蛋白EGFP-Prox1定位于PK15细胞的细胞核中。

Abstract:

Full length coding sequence of porcine Prox1 gene was cloned from liver cDNA. Then the chromosomal and subcellular localization of sProx1 were investigated. In this study, Blast searches was employed to obtain homologous porcine ESTs in the porcine EST databases with the Prox1 gene sequence of human and mouse, primers were designed and the mRNA sequence of Prox1 gene was amplified by RT-PCR. Blasted the intron2 sequence of Prox1 between human, mice and other kinds of species, a pair of primers was designed for the partly cloning of porcine Prox1 intron2. Chromosome localization of Prox1 was detected by IMpRH (INRA-Minnesota porcine radiation hybrid panel). pEGFP-N1-Prox1 was constructed and PK15 cells were transfected with the plasmid. The subcellular localization of Prox1 protein observed by inverted fluorescence microscope after 12 h. The complete Prox1 cDNA sequence and the intron2 partial sequence was cloned（GenBank Accession NO.EF486324，EF571585）. sProx1 gene was physically mapped on porcine chromosome 9q26 and closely linked to marker SW1651, and recombinant plasmid pEGFP-N1-Prox1 was constructed and EGFP-Prox1 was expressed in the nucleus of PK15 cells.

中图分类号:

S828
S813.3

袁继红，龙欢，田明芳，杨在清. 猪Prox1基因的染色体定位和亚细胞定位的研究[J]. 畜牧兽医学报, doi: .

YUAN Ji-hong, LONG Huan, TIAN Ming-fang, YANG Zai-qing. Study on Chromosomal and Subcellular Localization for Porcine Prox1 Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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