畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (9): 1371-1379.doi: 10.11843/j.issn.0366-6964.2013.09.006

• 遗传繁育 • 上一篇    下一篇

绵羊LPIN1基因的克隆和mRNA表达研究

魏琳琳1,高晋生2,王景霖1,程俐芬2,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*   

  1. (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)
  • 收稿日期:2013-05-08 出版日期:2013-09-23 发布日期:2013-09-23
  • 通讯作者: 刘文忠,E-mail: tglwzyc@163.com
  • 作者简介:魏琳琳(1987-),女,山西潞城人,硕士生,主要从事肉用绵羊的分子遗传育种研究,E-mail:dongkeweilin@163.com;高晋生(1962-),男,山西太原人,大学本科,高级畜牧师,主要从事肉用绵羊的分子遗传育种研究,E-mail:13753122191@139.com。二者为并列第一作者
  • 基金资助:

    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)

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

WEI Lin-lin1, GAO Jin-sheng2, WANG Jing-lin1, CHENG Li-fen2, QIAO Li-ying1, JIA Xia-li1, ZHANG Jing1, YANG Kai-jie1, LIU Jian-hua1, LIANG Chen1, LIU Wen-zhong1*   

  1. (1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China; 2. Shanxi Provincial Working Station for Animal and Poultry Breeding, Taiyuan 030001, China)
  • Received:2013-05-08 Online:2013-09-23 Published:2013-09-23

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2 688 bp,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。LPIN1 mRNA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。

Abstract:

This study aimed to clone ovine LPIN1 gene and to analyze its ontogenetic mRNA expression in seven adipose tissues of two breeds with distinct tail types, and based on which, to reveal 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 domains 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, retroperitoneal, 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 bp 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/Ned1/Smp2)).  LPIN1 mRNA expressed in all adipose tissues studied, but the highest in perirenal fat and the lowest in mesenteric 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 genetic improvement of meat quality traits of sheep based on its direct participating in and indirect regulating fat metabolism.

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