蒙古鮊ob基因的克隆与组织表达特异性的初步研究

根据已知的人和鼠ob基因序列设计引物,通过运用RT-PCR技术首次从蒙古鮊脂肪组织的RNA中扩增获得ob基因片段,并经序列分析证实为ob基因编码区438 bp的序列.不同物种同源性比较表明ob基因序列具有很高的保守性:蒙古鮊与人、猪和鼠的ob基因核苷酸编码序列的同源性分别为82.9%、84.0%和99.1%;蒙古鮊ob基因编码的蛋白leptin氨基酸序列与人、猪和鼠氨基酸同源性分别为80.8%、78.8%和95.9%.用RT-PCR技术分析组织ob基因的表达特异性,结果表明:ob基因在脂肪和肝脏组织中的表达量最大,在心脏、脾脏、肌肉、脑、卵巢中的表达量次之,在肾脏中仅微量表达,而在精巢和肠道组织中则不表达.

A Primary Study on the Expression of Cloning of ob Gene Cdna and Specific Tissue in Erythroculter mongolicus

In this study,the primers were designed according to the consensus region of human and mouse ob genes and segments of ob genes were been obtained from RNA tissue of Erythroculter mongolicus employing RT-PCR technique.Sequence analysis of these genes has confirmed that they belong to a 438bp sequence in coding region DNA.Congeneric analysis of different species shows that ob genes sequence has high conservative and progressive futures.Comparing with human,pig and mouse in the nucleotide sequences of ob genes,the homology of ob genes of Erythroculter mongolicus was 82.9%,84.0%,and 99.1% respectively.In comparison with human,pig and mouse in the Leptin amino acid sequence,the homology of amino acid sequences of Erythroculter mongolicus were 80.8%,78.8%,95.9% respectively.The result of ob gene tissue-specific expression by RT-PCR method indicates that ob gene was expressed mainly in adipose and liver,less in heart,spleen,muscle,brain and ovary,slightly in kidney,and none in spermary and intestines in E.mongolicus.