绵羊LPIN1基因的克隆和mRNA表达研究  

Study on the Cloning and Ontogenetic mRNA Expression of Ovine LPIN1 Gene

作　　者：魏琳琳[1] 高晋生 王景霖[1] 程俐芬 乔利英[1] 贾夏丽[1] 张静[1] 杨凯捷[1] 刘建华[1] 梁琛[1] 刘文忠[1] 

机构地区：[1]山西农业大学动物科技学院,太谷030801 [2]山西省畜禽繁育工作站,太原030001

出　　处：《畜牧兽医学报》2013年第9期1371-1379,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30972084;31372292);山西省科技攻关项目(011029);山西省研究生优秀创新项目(20133060)

摘　　要：本研究旨在克隆绵羊的LPIN1基因,分析其mRNA在2个不同脂尾型绵羊品种的7种脂肪组织中的发育性表达规律,以揭示该基因在绵羊脂肪代谢中的作用。利用RT-PCR技术克隆LPIN1基因,生物信息学方法分析Lipin1蛋白的理化性质和结构域;利用Real-time PCR技术检测广灵大尾羊和小尾寒羊2、4、6、8、10和12月龄共96个个体(每品种公、母各半)在大网膜、小网膜、肠系膜、腹膜后、皮下、肾周和尾部脂肪中的mRNA表达。结果表明,LPIN1基因CDS区长2 688bp,编码985个氨基酸。Lipin1是1种不稳定的亲水性蛋白,含有10个潜在的糖基化位点和88个磷酸化位点,但无跨膜域和信号肽。在其氨基酸序列的氨基端和羧基端各含有一段高度保守的序列,分别为Lipin-N和LNS2(Lipin/Ned1/Smp2)超家族特征序列。LPIN1mRNA在所研究的脂肪组织中都表达,其中肾周脂肪中表达最高,肠系膜脂肪中表达最低。品种、组织和月龄及其二阶互作对该基因的表达均有显著影响。LPIN1基因直接参与并间接调节脂肪代谢,对绵羊肉质性状的遗传改良具有重要意义。This study aimed to clone ovine LPIN1 gene and to analyze its ontogenetic mRNA ex- pression in seven adipose tissues of two breeds with distinct tail types, and based on which, to re- veal the role of the gene in fat metabolism of sheep. RT-PCR technique was used to amplify LPIN1 gene. Bioinformatics methods were used to predict the physicochemical properties and do- mains in Lipin1 protein. Ninety six animals from Guangling Large Tailed and Small Tailed Han （48 for each breed with equal sex ratios） were slaughtered at 2, 4, 6, 8, 10 and 12 months of age, respectively. Adipose tissues were collected from great and small omental, mesenteric, ret- roperitoneal, subcutaneous, perirenal and tail fats to study the ontogenetic mRNA expression by real-time PCR. The results showed that the coding region was 2 688 hp in length, coding 895 amino acids. Lipin1 was a kind of unstable and hydrophilic protein with 10 potential glycosylation and 88 phosphorylation sites, but without transmembrane domain and signal peptide. N- and C- terminals contained two highly conserved regions （Lipin-N and LNS2 （Lipin/Nedl/Smp2））.LPIN1 mRNA expressed in all adipose tissues studied, but the highest in perirenal fat and the lowest in mesenterie fats. Months of age, tissue and breed as well as their two-way interactions had significant influences on the LPIN1 mRNA expression. LPIN1 gene is significant in the ge- netic improvement of meat quality traits of sheep based on its direct participating in and indirect regulating fat metabolism.

关 键 词：绵羊 LPIN1基因 序列特征 脂肪组织 MRNA表达 

分 类 号：S826[农业科学—畜牧学] S813.3[农业科学—畜牧兽医]