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Tissue expression analysis, cloning and characterization of the 5'-regulatory region of the bovine FABP3 gene

机译:牛FABP3基因5'-调控区的组织表达分析,克隆与鉴定

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摘要

Fatty acid binding protein 3 (FABP3) is a member of the FABP family which bind fatty acids and have an important role in fatty acid metabolism. A large number of studies have shown that the genetic polymorphisms of FABP3 are positively correlated with intramuscular fat (IMF) content in domestic animals, however, the function and transcriptional characteristics of FABP3 in cattle remain unclear. Real-time PCR analysis revealed that bovine FABP3 was highly expressed in cardiac tissue. The 5'-regulatory region of bovine FABP3 was cloned and its transcription initiation sites were identified. Sequence analysis showed that many transcriptional factor binding sites including TATA-box and CCAAT-box were present on the 5'-flanking region of bovine FABP3, and four CpG islands were found on nucleotides from -891 to +118. Seven serial deletion constructs of the 5'-regulatory region evaluated in dual-luciferase reporter assay indicated that its core promoter was 384 base pairs upstream from the transcription initiation site. The transcriptional factor binding sites RXR alpha, KLF15, CREB and Sp1 were conserved in the core promoter of cattle, sheep, pigs and dogs. These results provide further understanding of the function and regulation mechanism of bovine FABP3.
机译:脂肪酸结合蛋白3(FABP3)是FABP家族的一个成员,可结合脂肪酸并在脂肪酸代谢中起重要作用。大量研究表明,FABP3的遗传多态性与家畜中的肌内脂肪(IMF)含量呈正相关,但是,牛中FABP3的功能和转录特性仍不清楚。实时PCR分析显示,牛FABP3在心脏组织中高度表达。克隆了牛FABP3的5'调节区,并鉴定了其转录起始位点。序列分析表明,在牛FABP3的5'侧翼区域存在许多转录因子结合位点,包括TATA-box和CCAAT-box,并且在-891至+118的核苷酸上发现了四个CpG岛。在双荧光素酶报告基因分析中评估的5'-调节区的七个序列缺失构建体表明,其核心启动子是转录起始位点上游的384个碱基对。转录因子结合位点RXR alpha,KLF15,CREB和Sp1在牛,绵羊,猪和狗的核心启动子中是保守的。这些结果提供了对牛FABP3的功能和调节机制的进一步理解。

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