多浪羊POMC基因克隆及其在初情期不同组织中的表达研究

Cloning of POMC Gene and Expression in Different Tissues of Duolang Sheep During Puberty

【目的】研究POMC基因在绵羊初情期时不同组织的表达水平及规律。【方法】对多浪羊进行初情期发情鉴定,采集初情期下丘脑、垂体、卵巢、输卵管、子宫、脾、肺、肾和大脑9种组织,提取其组织RNA,对POMC基因进行cDNA克隆和组织表达分析;对多浪羊POMC氨基酸与其他物种POMC氨基酸序列进行比较,制作系统发育树;应用qPCR技术检测POMC基因在下丘脑、垂体、卵巢、输卵管和子宫5种组织中的表达水平。【结果】获得多浪羊POMC基因两条cDNA序列,长度分别为1 078、1 108 bp,两条cDNA序列编码区(CDS)相同,均为792 bp,编码263个氨基酸。同源性比对分析得出多浪羊POMC氨基酸序列与绵羊、牛、山羊的同源性分别为99.62%、96.6%、96.3%,与猪、马、人的同源性分别为89.51%、83.08%、79.78%。qPCR和组织表达谱分析结果表明POMC基因在9种组织均有表达,其中在垂体和下丘脑中表达量比较高,显著高于其他组织(P<0.05)。【结论】试验结果对于进一步探索POMC基因与绵羊初情期之间的关系提供了理论依据。

【Objective】 In order to study the expression level of the POMC gene in various tissues of Duolang sheep during puberty.【Method】In this experiment,Duolang sheep was used as the experimental material,at first,the puberty identification was carried out. Nine tissues of hypothalamus,pituitary gland,ovary,fallopian tube,uterus,spleen,lung,kidney and brain were collected,the RNA of the tissues was extracted,the gene cloning and tissue expression profile analysis of the Duolang sheep POMC gene were carried out,the identity of the aminoacid sequences between Duolang sheep and other species was compared,then the phylogenetic tree was constructed,the expression level of the POMC gene in the five tissues of the hypothalamus,pituitary,ovary,uterus and fallopian tubewas detected by qPCR.【Result】Two cDNA sequences of POMC gene of Duolang sheep were obtained,the length of the first cDNA sequence was 1 078 bp,and the second cDNA sequence was 1 108 bp. The coding region(CDS)of the two c DNA sequences was the same,both of which were 792 bp,encoding 263 amino acids.The homology comparison analysis showed that the amino acid sequence of Duolang sheep POMC was 99.62%,96.6%,and 96.3% with those of sheep,cattle,and goats,respectively. The homology with pigs,horses,and humans was 89.51%,83.08%,and 79.78%,respectively. The results of qPCR and tissue expression profiling showed that the POMC gene was expressed in the mentioned tissues,among which it was significantly expressed in the pituitary and hypothalamus(P<0.05).【Conclusion】The results of this experiment provided a scientific theoretical basis for further exploring the relationship between the POMC gene and the puberty of sheep.