山羊DCT基因启动子活性区及其转录因子调控探究

doi:10.11843/j.issn.0366-6964.2018.01.007

Abstract

Abstract:

The research aimed to study the promoter activity area and to explore the regulation mechanism of some related transcription factors of goat DCT gene, which would help to find the theoretical reference for expression regulation of goat DCT. 5' flanking region and the first exon sequences were analyzed by bioinformatics and blasted with mouse and human DCT gene promoter sequences. Based on the results of bioinformatics analysis and online promoter prediction, five 5'-terminal and six 3'-terminal deleted fragment promoter reporter gene vectors were constructed with the rapid amplification kit. Six mutation reporter gene vectors for SOX10, MITF and OTX2 transcription factor binding sites were built using P8 fragment sequences as the template. These vectors were transfected into A375 cells by transient transfection and their promoter activities were detected with the dual-luciferase detection reagent. The results showed that 11 deleted fragment promoter reporter gene vectors with different length were successfully constructed. The luciferase activity of P3 vector for -990-+232 bp was significantly higher than the others (P<0.01). And P8 vector for -881——154 bp based on P3 fragment had the highest luciferase activity among the serial 3'-terminal deletion vectors (P<0.01). The mutation vectors of SOX10 binding site showed extremely significantly lower luciferase activity compared with the original vector (P<0.01), while the MITF and OTX2 binding sites mutation vectors both showed extremely significantly higher luciferase activity compared with the original vectors (P<0.01). The core promoter region of goat DCT gene is located at -881——154 bp. Transcription factor SOX10 up-regulate the expression of DCT gene, while the regulating role of transcription factor MITF and OTX2 should be thoroughly studied.

CLC Number:

S827.2

LIU Chun-yang, ZHANG Le-chao, WANG Qi, ZHOU Rong-yan, LI Lan-hui, LI Xiang-long. The Exploration of the Promoter Activity Area and Regulation by Transcription Factors of Goat DCT Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 55-64.

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The Exploration of the Promoter Activity Area and Regulation by Transcription Factors of Goat DCT Gene

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