绵羊脂肪细胞型脂肪酸结合蛋白基因(FABP4)cDNA的克隆、表达及其结构模拟分析  

Cloning,Expression and Structure Modeling Analysis of Adipocyte Fatty Acid Binding Protein Gene(FABP4) of Sheep

作　　者：徐秋良[1,2] 张庆莉[2] 陈玉林[1] 

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100 [2]郑州牧业工程高等专科学校畜牧工程系,郑州450011

出　　处：《农业生物技术学报》2011年第3期483-489,共7页Journal of Agricultural Biotechnology

基　　金：国家重大转基因专项"高产绒量转基因绒山羊新品种培育"(No.2008ZX08008-002);国家绒毛用羊产业技术体系(No.nycytx-40-11);郑州牧业工程高等专科学校学术带头人基金共同资助

摘　　要：为探讨绵羊脂肪细胞型脂肪酸结合蛋白(FABP4)在不同生长阶段背最长肌中的表达特性,本实验以成年小尾寒羊(Ovisaries)皮下脂肪组织总RNA为实验材料,在脂肪细胞型脂肪酸结合蛋白基因(FABP4)cDNA保守区域设计1对引物Ff和Fr,采用RT-PCR方法克隆了绵羊FABP4全编码序列,并运用生物软件对FABP4蛋白空间结构进行分析。同时运用实时荧光定量PCR法,对FABP4在皮下脂肪和背最长肌中的表达以及不同日龄在背最长肌中的表达量与背最长肌肌内脂肪含量的相关性进行了初步研究。结果表明:绵羊FABP4基因cDNA长度为464bp,包含1个由399bp组成的开放阅读框,编码132个氨基酸。生物信息学分析表明,不同物种间FABP4氨基酸序列具有较强的保守性,蛋白质空间结构呈2个α螺旋和10个β折叠围绕的桶状构型。实时荧光定量PCR试验表明,FABP4基因在160和200日龄绵羊背最长肌中的表达量明显高于90日龄背最长肌的表达量(P<0.05);且在90、120、160和200日龄绵羊背最长肌中的表达量与相应日龄的背最长肌肌内脂肪含量呈正相关(R2=0.7196,P<0.01)。本研究结果表明,调控FABP4基因在背最长肌的表达是改善绵羊肉质的潜在途径。For investigated the expression level of adipocyte fatty acid binding protein gene（FABP4） at different stages of sheep longissimus muscle,in this paper,the cDNA of sheep（Ovis aries） adipocyte fatty acid binding protein gene（FABP4） was cloned by the method of RT-PCR using designed primer pairs of Ff and Fr according to the FABP4 gene cDNA conservative region.The putative protein spatial structure of FABP4 was analyzed by biology software.Additionally,the expression of FABP4 in sheep longissimus muscle and the correlation between FABP4 expressions of longissimus muscle and intramuscular fat contents at different days were tested by real time RT-PCR.The results showed that the sequence of cDNA of sheep FABP4 was 464 bp containing a 399 bp of open reading frame which encoded 132 amino acids and the amino acid of FABP4 was conserved in evolution,and the FABP4 protein was assembled as a barrel conformation by two α helixes and ten β folds.The real time RT-PCR revealed that expressions of FABP4 in longissimus muscle from 160 and 200-day＇s sheep were higher than those from 90-day＇s（P0.05）,respectively.The expression of FABP4 longissimus muscle at different days was positive correlated with longissimus muscle intramuscular fat contents（R2=0.7196,P0.01）.The result indicated that regulated the expression of FABP4 in sheep longissimus muscle is a potential approach for improving sheep meat quality traits.

关 键 词：脂肪细胞型脂肪酸结合蛋白基因FABP4 CDNA克隆 绵羊 表达 结构模拟 

分 类 号：S831.2[农业科学—畜牧学]