Tissue expression analysis, cloning and characterization of the 5'-regulatory region of the bovine FABP3 gene
Tissue expression analysis, cloning and characterization of the 5'-regulatory region of the bovine FABP3 gene
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DOI：
10.1007/s11033-016-4026-7
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发表时间：
2016-09-01
影响因子：
2.8
通讯作者：
Zan, Linsen
中科院分区：
文献类型：
--
作者：
Li, Anning;Wu, Lijuan;Zan, Linsen
脂肪酸结合蛋白3 （Fatty acid binding protein 3, FABP3）是与脂肪酸结合的FABP家族成员，在脂肪酸代谢中起重要作用。大量研究表明，家畜FABP3的遗传多态性与肌内脂肪（IMF）含量呈正相关，但牛FABP3的功能和转录特征尚不清楚。实时荧光定量PCR结果显示，牛FABP3在心脏组织中高表达。克隆了牛FABP3的5′调控区，并确定了其转录起始位点。序列分析表明，牛FABP3的5′侧区存在多个转录因子结合位点，包括TATA-box和CCAAT-box，在-891 ~ +118核苷酸上发现4个CpG岛。双荧光素酶报告基因分析显示，5'调控区的7个序列缺失结构表明，其核心启动子位于转录起始位点上游384个碱基对。转录因子结合位点RXR α、KLF15、CREB和Sp1保守存在于牛、羊、猪和狗的核心启动子中。这些结果为进一步了解牛FABP3的功能及其调控机制提供了依据。
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.