秦川牛A-FABP基因的生物信息学分析

Biological information analysis of A-FABP gene in Qinchuan cattle

【目的】克隆秦川牛脂肪型脂肪酸结合蛋白(A-FABP)基因,并对其进行生物信息学分析,为深入研究A-FABP基因与秦川牛肉质性状的关系奠定基础。【方法】以秦川牛脂肪组织为材料,采用RT-PCR方法对牛A-FABP基因进行克隆。使用DNAMAN、NCBI等一系列在线软件及工具,对所得到的序列及其编码蛋白质的结构和特性等进行生物信息学分析。【结果】秦川牛A-FABP基因含有15个酶切位点,编码的蛋白分子质量为14.7 ku,二级结构主要以α-螺旋、不规则盘绕和延伸链为结构元件,含132个氨基酸,其中强碱性氨基酸18个,强酸性氨基酸19个,疏水氨基酸45个,不带电荷的极性氨基酸32个;含有6个蛋白激酶C磷酸化位点,1个酪蛋白激酶Ⅱ磷酸化位点,3个肉豆蔻酰基化位点,1个FABP结合域;含有4个Ser、6个Thr和1个Tyres,这11个氨基酸均可能成为蛋白激酶磷酸化位点。【结论】秦川牛A-FABP基因编码的氨基酸序列与人、鸡、小鼠、大鼠、野猪的氨基酸序列同源性分别为84.09%,85.61%,71.97%,88.33%,81.82%,表明在进化关系上,A-FABP氨基酸序列有较好的保守性;此蛋白序列具有脂钙蛋白结合域,没有信号肽;无明显跨膜区,不含有二硫键。

【Objective】 A-FABP gene in Qinchuan cattle was cloned and its biological information was analyzed,which might provide more information to the further study of A-FABP gene.【Method】 This experiment extracted the total RNA from Qinchuan cattle fat tissue,and A-FABP gene CDS region was cloned by RT-PCR in pMD18-T vector,and then sequenced.The sequence was analyzed by DNAMAN,NCBI,and some other software and online tools in biological information.【Result】 The analysis of amino acid sequence indicated that Qinchuan cattle A-FABP gene included 15 restriction enzyme cutting sites and coded 132 amino acids,molecular weight 14.7 ku,main elements of its secondary structure being α-helix,random coil and extended strand,including 6 protein kinase C phosphorylation sites,1 casein kinase phosphorylation site,3 N-myristoylation sites and 1 cytosolic fatty-acid binding protein signature as well as 4 Ser,6 Thr and 1 Tyr,which could be protein kinase phosphorylation site.【Conclusion】 Homologous comparison with some animals indicated that Qinchuan cattle A-FABP shares 84.09%,85.61%,71.97%,88.33%,81.82% similarity in nucleic acid sequence with man,chicken,mouse,rats,and swines.Bioinformatics analysis indicated that A-FABP gene has high conservatism.