牛IRX3基因启动子转录调控分析

doi:10.11843/j.issn.0366-6964.2020.05.011

Abstract

Abstract: The aim of this study was to elucidate the transcriptional regulation mechanism and expression patterns of bovine IRX3 gene and identify the key transcription factors of promoter. The distribution of bovine IRX3 mRNA in 3 adult bulls tissues was determined through qPCR, including heart, liver, spleen, lung, kidney, subcutaneous fat, longissimus dorsi, large intestine, small intestine, reticulum, rumen, abdomen and testicle. In addition, the 1.8 kb promoter sequence of IRX3 gene was cloned and dual luciferase receptor vectors with the 6 different deletion fragments of the promoter were constructed, and then transfected into 3T3-L1 and C2C12 cell lines, respectively. Furthermore, the key transcription factors in the core promoter region were predicted by online software, and the transcriptional regulation of key transcription factors on IRX3 gene was identified in 3T3-L1 cell line by site-directed mutation and siRNA interference. The results showed that IRX3 had a broad tissue distribution in 13 tissues, and it was highly expressed in the lung, kidney, heart, subcutaneous fat and longissimus dorsi (P<0.05). The promoter core region of bovine IRX3 gene was located at -372/-42 bp using the dual luciferase reporter vector. Combined with site-directed mutation and siRNA interference demonstrated that NRF1, KLF4, HOXA5 and CREB1 regulated the transcriptional activity of IRX3 gene. The tissue expression analysis results indicated that the expression level of IRX3 was relatively higher in longissimus dorsi and subcutaneous fat. There were 8 CpG islands in the 1.8 kb region of IRX3 promoter. NRF1, KLF4, HOXA5 and CREB1 were located in CpG islands and regulated the transcriptional activity of IRX3 gene. These results lay an important theoretical foundation for exploring the molecular regulation mechanism of IRX3 gene in bovine fat deposition.

Key words: IRX3 gene, promoter, transcriptional regulation

CLC Number:

S823.2

WEI Dawei, YANG Zhiyan, WANG Xingping, LUO Rengzhuoma, ZHANG Jiupan. Transcription Regulation Analysis of Bovine IRX3 Gene Promoter[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 997-1006.

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Transcription Regulation Analysis of Bovine IRX3 Gene Promoter

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