中国农业科学2013，Vol.46Issue(10)：2110-2118,9.DOI:10.3864/j.issn.0578-1752.2013.10.017
绵羊UCP4基因编码序列的克隆和mRNA表达研究
Study on the Cloning of Coding Sequence and Ontogenetic mRNA Expression of Ovine UCP4 Gene
摘要
Abstract
[Objective] This study aims to clone the coding sequence (CDS) of ovine UCP4 gene,to analyze the CDS and its coding protein structures,and to profile the ontogenetic mRNA expression so that to lay a theoretical foundation for future research on the structure and function of this gene.[Method] The CDS was amplified from ovine cerebrum tissue by reverse transcriptionpolymerase chain reaction (RT-PCR).Physicochemical features and structural characteristics of ovine UCP4 were analyzed by bioinformatics methods.Ninety-six animals from two sheep breeds with significant differences in tail types (Guangling Large Tailedand Small Tailed-Han Sheep) were used to study the ontogenetic mRNA expression by real-time quantitative PCR in eight tissues (cerebrum,cerebellum,hypothalamus,pituitary,and subcutaneous fat,perirenal fat,mesenteric fat and tail fat) from 2-to 12-month old age at two-month interval.[Result] The sequencing data showed that the length of complete CDS was 972 bp encoding a protein of 323 amino acids.The isoelectric point (pI) of the protein was 9.43,and the molecular weight was 35.73 kDa.The percentages of helix,strand and loop were 56.04％,7.12％ and 36.84％,respectively,in the secondary structure.This transmembrane protein had no signal peptide but with two predicted glycosylation sites and 15 potential phosphorylation sites.The UCP4 mRNA expressed in all these tissues,but overexpressed in brain tissue other than in adipose tissue.Generally,breed,tissue and the months of age had significant influences on the UCP4 mRNA expression.[Conclusion] The obtained whole CDS of ovine UCP4 gene,expression profile and its limiting factors are of scientific significance to further studies on the relationship between the gene structure and energy metabolism.关键词
绵羊/UCP4基因/编码序列/mRNA表达Key words
sheep/ UCP4 gene/ coding sequence/ mRNA expression引用本文复制引用
周沙沙,刘宝凤,魏琳琳,王景霖,刘建华,梁琛,乔利英,刘文忠..绵羊UCP4基因编码序列的克隆和mRNA表达研究[J].中国农业科学,2013,46(10):2110-2118,9.基金项目
国家自然科学基金(30972084)、山西省科技攻关项目(011029)、山西农业大学中青年拔尖创新人才项目(BJRC201203) （30972084）
