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Journal of Zhejiang University SCIENCE B

ISSN 1673-1581(Print), 1862-1783(Online), Monthly

Molecular cloning, characterization and expression of the energy homeostasis-associated gene in piglet

Abstract: The energy homeostasis-associated (Enho) gene encodes a secreted protein, adropin, which regulates the expression of hepatic lipogenic genes and adipose tissue peroxisome proliferator-activated receptor γ, a major regulator of lipogenesis. In the present study, the porcine (Sus scrofa) homologue of the Enho gene, which was named pEnho, was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) using oligonucleotide primers derived from in silico sequences. The gene sequence was submitted into the GenBank of NCBI, and the access number is GQ414763. The pEnho encodes a protein of 76 amino acids which shows 75% similarity to Homo sapiens adropin. The expression profile of pEnho in tissues (liver, muscle, anterior jejunum, posterior jejunum, and ileum) was determined by quantitative real-time RT-PCR. pEnho was localized on porcine chromosome 10 and no introns were found. In conclusion, pEnho was cloned and analysed with the aim of increasing knowledge about glucose and lipid metabolism in piglets and helping to promote the health and growth of piglets through adropin regulation.

Key words: Adropin, Energy homeostasis-associated (Enho) gene, Gene coloning, Piglets, RT-PCR

Chinese Summary  <26> 猪Enho基因的分子克隆及其仔猪的组织分布

目的:通过pEnho基因的克隆,分析pEnho基因在仔猪相关组织的分布情况,增加仔猪糖脂代谢方面相关的知识,为通过调控adropin来促进仔猪的健康和生长提供理论基础。
创新点:首次克隆了猪Enho基因,命名为pEnho,其基因序列提交至美国国立生物技术信息中心(NCBI)(No.GQ414763)。
方法:通过实时荧光定量逆转录聚合酶链式反应(real-timeRT-PCR)方法获得猪Enho基因序列,并应用BLAST、ClustalW、PHYLIP、TMHMM、Helixturnhelix等生物信息学软件对其基因序列进行分析;通过荧光定量PCR方法检测了不同日龄(出生后1、7、14和21天)猪Enho基因的组织分布(肝脏、肌肉、空肠前端、空肠后端和回肠)。
结论:克隆得到pEnho基因序列;发现随着仔猪日龄的增加,其表达量有下降;在组织分布上,产后7天内pEnho在肠道(特别是回肠)的表达量较高,在第14天和第21天,各被检组织的表达量无显著差异。

关键词组:Adropin;Enho;基因克隆;仔猪;定量PCR


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DOI:

10.1631/jzus.B1400260

CLC number:

S828

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On-line Access:

2015-06-08

Received:

2014-09-26

Revision Accepted:

2015-03-12

Crosschecked:

2015-05-13

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