八眉猪FABPs主要家族基因单核苷酸多态性筛查及生物信息学分析

Single nucleotide polymorphism screening and bioinformatics analysis of the main family genes of FABPs in Bamei pig

脂肪酸结合蛋白(fatty acid-binding protein, FABPs)属细胞内脂质结合蛋白超家族,FABPs家族所有成员都具有管理游离脂肪酸吸收和细胞内转运的最基本功能。为了探究FABPs主要家族基因在八眉猪体内脂肪沉积代谢过程中的调控机制,本研究以八眉猪为试验对象构建DNA混合池,采用分段扩增和直接测序法对八眉猪FABP_1、FABP_2、FABP_3、FABP_4基因进行单核苷酸多态性(single nucleotide polymorphisms, SNPs)快速筛查和生物信息学分析。结果表明,在八眉猪FABPs主要家族基因中共检测出15个SNPs,其中编码区有3个同义突变和2个错义突变(C^(2033)A-Exon2-FABP_1、G^(3 751)A-Exon3-FABP_1)。对其进行生物信息学分析表明:核苷酸突变前后mRNA二级结构和自由能的改变导致其结构稳定性发生改变;突变前后的等位基因频率估算,mRNA二级结构预测,蛋白质二、三级结构预测分析均有差异。说明八眉猪FABPs主要家族基因具有较高的突变性,为研究八眉猪肌内脂肪含量的遗传效应提供了分子理论依据。

Fatty acid-binding proteins belong to the intracellular lipid binding protein superfamily. All members of the FABPs family have the most basic functions to manage free fatty acid absorption and intracellular transport. In order to explore the regulation mechanism of the FABPs main family genes on the lipid deposition and metabolism in Bamei pigs, in this study, DNA pooling was constructed with Bamei pigs as test objects, and the single nucleotide polymorphisms(SNPs) of FABP1, FABP2, FABP3 and FABP4 genes in Bamei pigs were screened by segmented amplification and direct sequencing, and the bioinformatics analysis was carried out. The result showed that 15 SNPs were detected in the FABPs main family genes, in which there were three samesense and two missense mutation(C2 033A-Exon2-FABP1, G3 751A-Exon3-FABP1). Bioinformatics analysis showed that the secondary structure of mRNA and free energy change before and after nucleotide mutation led to the change of its structural stability;there were differences in the estimation of allele frequency, mRNA secondary structure prediction, and protein secondary and tertiary structure prediction before and after the mutation. The above results indicate that the FABPs main family genes have high mutagenicity, which provides a molecular basis for the genetic effect study of intramuscular fat content in Bamei pigs.