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一种在梅山猪、梅山猪与大白猪杂交猪以及大白猪的背最长肌组织中差异表达的新型猪基因CXCL10的分离、序列分析及表达谱研究

Isolation, sequence analysis and expression profile of a novel porcine gene, CXCL10, differentially expressed in the Longissimus dorsi muscle tissues from Meishan, Meishan x Large White cross and Large White pigs.

作者信息

Liu G Y, Xiong Y Z

机构信息

Key Laboratory of Animal Nutrition and Feed of Yunnan Province, Yunnan Agricultural University, Kunming, People's Republic of China.

出版信息

DNA Seq. 2007 Dec;18(6):415-22. doi: 10.1080/10425170701243328.

Abstract

In order to detect the molecular mechanism of heterosis in pigs, the mRNA differential display (DD) technique was performed to investigate the differences in gene expression in the Longissimus dorsi muscle tissues from Meishan (MS), Meishan x Large White (ML) cross and Large White (LW) pigs. One novel gene that was differentially expressed was identified using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and its full-length cDNA sequence was obtained using the rapid amplification of cDNA ends (RACE) method. The nucleotide sequence of the gene is not homologous to any of the known porcine genes. The sequence prediction analysis revealed that the open reading frame of this gene encodes a protein of 104 amino acids that contains the putative conserved domain of the chemokine CXC which could be designated as chemokine cd00273 subgroup and this protein has high homology with the small inducible cytokine B10 precursor (CXCL10) of five species - dog(87%), human (84%), monkey (84%), mouse (75%) and rat (70%) - so that it can be defined as swine small inducible cytokine B10 precursor. The phylogenetic tree analysis revealed that the swine CXCL10 has a closer genetic relationship with the CXCL10 of dog than with those of human, monkey, mouse and rat. The tissue expression analysis indicated that the swine CXCL10 gene is more highly expressed in muscle and weakly expressed in fat and kidney. The genomic sequence of swine CXCL10 gene was finally amplified and result revealed that the swine CXCL10 gene contains four exons and three introns. Our experiment is the first to establish the primary foundation for further research on the swine CXCL10 gene.

摘要

为了探究猪杂种优势的分子机制,运用mRNA差异显示(DD)技术,研究梅山猪(MS)、梅山猪与大白猪杂交后代(ML)以及大白猪(LW)背最长肌组织中的基因表达差异。通过半定量逆转录聚合酶链反应(RT-PCR)鉴定出一个差异表达的新基因,并利用cDNA末端快速扩增(RACE)方法获得其全长cDNA序列。该基因的核苷酸序列与任何已知猪基因均无同源性。序列预测分析表明,该基因的开放阅读框编码一个含104个氨基酸的蛋白质,该蛋白质含有趋化因子CXC的假定保守结构域,可归为趋化因子cd00273亚组,且与犬(87%)、人(84%)、猴(84%)、小鼠(75%)和大鼠(70%)这五个物种的小诱导细胞因子B10前体(CXCL10)具有高度同源性,因此可定义为猪小诱导细胞因子B10前体。系统发育树分析显示,猪CXCL10与犬的CXCL10的亲缘关系比与人类、猴、小鼠和大鼠更为密切。组织表达分析表明,猪CXCL10基因在肌肉中表达较高,在脂肪和肾脏中表达较弱。最终扩增出猪CXCL10基因的基因组序列,结果表明该基因含有四个外显子和三个内含子。我们的实验首次为进一步研究猪CXCL10基因奠定了初步基础。

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