Liu G Y, Xiong Y Z
Yunnan Agricultural University, Kunming, 650201, China.
Mol Biol Rep. 2009 Jan;36(1):57-62. doi: 10.1007/s11033-007-9151-x. Epub 2007 Oct 13.
In order to detect the molecular mechanism of heterosis in pigs, the mRNA differential display technique was performed to investigate the differences in gene expression in the longissimus dorsi muscle tissues from Meishan, Meishan x Large White cross and Large White 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. Sequence analysis revealed that open reading frame of this gene encoding a protein of 105 amid acids and this protein showed 100% homology to bovine and ovine CYCS, and therefore, this gene can be defined as the swine CYCS gene. The genomic sequence of swine CYCS gene was finally amplified and result revealed that the swine CYCS gene contains no introns. Tissue expression profile analysis revealed that swine CYCS gene was highly expressed in muscle, fat and lung, moderately expressed in ovary, kidney, and liver, and weekly expressed in heart, spleen and small intestine. Our results established the primary foundation for further research into the biological significance of swine CYCS gene.
为了探究猪杂种优势的分子机制,采用mRNA差异显示技术,研究梅山猪、梅山猪与大白猪杂交后代以及大白猪背最长肌组织中的基因表达差异。利用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,并通过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。序列分析表明,该基因的开放阅读框编码一个含105个氨基酸的蛋白质,该蛋白质与牛和羊的CYCS具有100%的同源性,因此,该基因可定义为猪CYCS基因。最终扩增出猪CYCS基因的基因组序列,结果显示该基因不含内含子。组织表达谱分析表明,猪CYCS基因在肌肉、脂肪和肺中高表达,在卵巢、肾脏和肝脏中中度表达,在心脏、脾脏和小肠中低表达。我们的研究结果为进一步研究猪CYCS基因的生物学意义奠定了初步基础。