牛GPAM基因分子克隆及编码区E1-76-C>T定点突变前后生物信息学对比分析

Cloning of Bovine GPAM Gene and Comparative Analysis of Bioinformatics Before and After Coding Region E1- 76- C > T Site-Specific Mutagenesis

线粒体甘油-3-磷酸酰基转移酶基因(Mitochondrial glycerol-3-phosphate acyltransferase,GPAM)的主要作用是通过催化三酰基甘油和磷脂生物合成过程,从而促进动物机体甘油三酯的生成,并通过中心和外周调节作用,对哺乳动物脂肪沉积、能量消耗、胴体重以及整体的代谢进行调控。设计特定引物对牛GPAM基因编码区进行PCR扩增并进行了克隆及测序,并假定牛GPAM基因编码序列发生E1-76-C>T点突变,通过生物信息学的方法,对牛GPAM基因突变前后编码产物的理化性质、结构与功能进行了对比分析,通过生物信息学对比分析得出,牛GPAM基因单碱基的改变可以影响到氨基酸的编码,在一级结构、二级结构和三级结构方面存在显著差异。

The main function of Mitochondrial glycerol-3-phosphate acyltransferase（ GPAM） gene is to promote production of triglyceride in animal body through catalytic triacylglycero phospholipid bio-synthesis, and to regulate fat deposition, energy consumption, carcass weight and overall metabolism of mammals through central and peripheral regulating action. In this study, coding region of bovine GPAM gene was amplified via specific primers by PCR method, and was cloned and sequenced. Bioinformatics methods were used for comparatively analyzing physical and chemical properties, structure and function between mutant type and normal type, assuming that mutation occurred withE1-76-C〉T site of bovine GPAM gene coding sequence. Compartive analysis of bioinformatics showed that bovine GPAM gene single nucleotide change can affect amino acid coding, and there are significant differences in primary structure, secondary structure and tertiary structure of protein.