【摘要】 本研究以湖羊肝脏为材料对肝受体类似物-1(Liver receptor homolog-1,Lrh-1;NR5A2)基因序列进行RT-PCR和RACE测定,并用DNAman、Tmpred、Signal P 3.0、Expasy等生物信息学分析软件和在线工具,对Lrh-1cDNA序列及其蛋白的理化特性、跨膜结构、信号肽和二级结构进行生物信息学分析,同时采用实时荧光定量PCR方法检测湖羊Lrh-1基因的组织表达谱。结果表明,湖羊Lrh-1基因cDNA序列全长1 488bp,与牛的核酸序列相似度最高,为98%,编码区共编码495个氨基酸,氨基酸序列与牛、人、马、猴、犬、小鼠、褐鼠的氨基酸同源性分别为98%、97%、96%、97%、97%、86%和86%;Lrh-1mRNA在湖羊脑、消化及生殖系统组织中均有表达且具有组织特异性,其中在下丘脑组织中的表达量相对最高。更多还原

【Abstract】 In the study,the full-length cDNA of the liver receptor homolog-1(Lrh-1) was detected from liver of Hu sheep by reverse transcription PCR and RACE methods.Lrh-1 cDNA sequence,protein physicochemical properties,membrane structure,signal peptide and secondary structural characterisitics were analyzed and predicted with bioinformatics softwares and online tools,such as DNAman,Tmpred,SignalP 3.0,TargetP 1.1,Expasy,and PSORT II prediction.The expression of Lrh-1 mRNA in different tissues of Hu sheep was also examined by quantitative real-time PCR.The results showed that the cDNA of Hu sheep Lrh-1 contained 1 488 nucleotides,and the nucleotide sequence had 98% homology with that of Bos taurus.It encoded 495 amino acid residues,which shared 98%,97%,96%,97%,97%,86% and 86% identity with those of Bos taurus,Homo sapiens,Equus caballus,Macaca mulatta,Canis familiaris,Mus musculus and Rattus norvegicus,respectively.The relative expression level of Lrh-1 mRNA was detected in brain,digestive system and reproductive system,and the expression levels were showed differently and had tissue-specific.Relatively,Lrh-1 mRNA expression level was the highest in hypothalamus of Hu sheep.更多还原