贵州白山羊MyoG基因启动子区序列克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2023.03.005

摘要/Abstract

摘要： 【目的】本研究旨在解析肌细胞生成素(MyoG)基因启动子区序列的结构特性及其转录调控机制。【方法】以贵州白山羊血液基因组DNA为模板,设计2对引物,采用PCR扩增、Sanger测序技术获得MyoG基因启动子区序列,通过DNAStar和Mega 5.0软件分别进行不同物种MyoG基因启动子区序列相似性比对及系统进化树构建,利用生物信息学软件预测分析该序列核心启动子区、转录因子结合位点、顺式作用元件、CpG岛等结构特征。【结果】贵州白山羊MyoG基因启动子区序列长2334 bp,包括2092 bp的5'-侧翼序列和242 bp的外显子1,GC含量略低于AT含量。通过不同物种比对分析,贵州白山羊MyoG基因启动子区序列与绵羊、牛、猪、人、小鼠和鸡同源序列相似性分别为98.34%、96.15%、77.76%、74.15%、57.63%和43.04%。系统进化树结果表明,贵州白山羊与绵羊和牛的亲缘关系较近,与鸡的亲缘关系最远。生物信息学结构预测发现,贵州白山羊MyoG基因转录起始位点位于翻译起始密码子ATG上游50 bp,5'-侧翼区存在1个潜在的启动子区域和1个CpG岛,含有1个TATA-box(TTTAAATG)、1个GC-box(CCGCCC)、2个CAAT-box(CCAAT)、17个E-box(CANNTG)和1个富含A/T碱基的元件结构(TATATTTAT)。AliBaba 2.1、PROMO、Patch 1.0、JASPAR²⁰²²和AnimalTFDB 3.0在线软件综合预测发现,贵州白山羊MyoG基因启动子区存在Sp1、NF-1、MEF2、MyoD、USF、GATA-1、GATA-3、SRF、C/EBP和c-Myb等多种转录因子潜在结合位点。【结论】本研究成功克隆获得贵州白山羊MyoG基因启动子区序列,为深入解析该基因调控山羊肌肉发育的分子机制提供理论依据。

关键词: 贵州白山羊; MyoG基因; 启动子区; 转录因子; 生物信息学

Abstract: 【Objective】 The purpose of this study was to analyze the structural characteristics of the promoter region sequence of MyoG gene and its transcriptional regulation mechanism.【Method】 The blood genomic DNA of Guizhou White goat was used as template,two pairs of primers were designed,and the promoter region of MyoG gene was obtained by PCR amplification and Sanger sequencing.The sequence similarity comparison and phylogenetic tree construction of MyoG gene promoter region among different species were carried out by DNAStar and Mega 5.0 software.The core promoter region,transcription factor binding site,cis-acting element and CpG island of obtained sequence were predicted and analyzed by bioinformatics software.【Result】 The length of promoter region of MyoG gene in Guizhou White goat was 2 334 bp,including the 5'-flanking region of 2 092 bp and 242 bp of exon 1,and the GC content was slightly lower than that of AT.Through the comparative analysis of different species,the similarity of the promoter region of MyoG gene in Guizhou White goat with that of Ovis aries,Bos taurus,Sus scrofa,Homo sapiens,Mus musculus,and Gallus gallus were 98.34%,96.15%,77.76%,74.15%,57.63% and 43.04%,respectively.Phylogenetic tree revealed that Guizhou White goat was the most closely related to Ovis aries and Bos taurus,and the farest related was Gallus gallus.Bioinformatics structure prediction found that the transcription initiation site of MyoG gene in Guizhou White goat was located at 50 bp upstream of the translation initiation codon ATG,and there was a potential promoter region and a CpG island in the 5'-flanking region,containing 1 TATA-box (TTTAAATG),1 GC-box(CCGCCC),2 CAAT-box(CCAAT),17 E-box(CAGATG) and 1 A/T rich element (TATATTTAT). The potential binding sites of Sp1,NF-1,MEF2,MyoD,USF,GATA-1,GATA-3,SRF,C/EBP and c-Myb transcription factors were predicted by AliBaba 2.1,PROMO,Patch 1.0,JASPAR²⁰²² and AnimalTFDB 3.0 online software.【Conclusion】 The promoter region sequence of MyoG gene in Guizhou White goat was successfully cloned,which provided a theoretical basis for further research on the molecular mechanism of MyoG gene regulating goat muscle development.

Key words: Guizhou White goat; MyoG gene; promoter region; transcription factor; bioinformatics

中图分类号:

S827

宋兴超, 孟金柱, 赵园园, 吴震洋, 安清明. 贵州白山羊MyoG基因启动子区序列克隆及生物信息学分析[J]. 中国畜牧兽医, 2023, 50(3): 893-903.

SONG Xingchao, MENG Jinzhu, ZHAO Yuanyuan, WU Zhenyang, AN Qingming. Cloning and Bioinformatics Analysis of MyoG Gene Promoter Region Sequence in Guizhou White Goat[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 893-903.

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