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Cloning and Bioinformatics Analysis of MyoG Gene Promoter Region Sequence in Guizhou White Goat
- SONG Xingchao, MENG Jinzhu, ZHAO Yuanyuan, WU Zhenyang, AN Qingming
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2023, 50(3):
893-903.
doi:10.16431/j.cnki.1671-7236.2023.03.005
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【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.