海南黑山羊MBL2基因的转录调控元件的筛选与分析

doi:10.11843/j.issn.0366-6964.2022.06.013

Abstract

Abstract: Mannose binding lectin C(MBL-C)is a member of the C-type (Ca²⁺dependent) lectin superfamily. As an acute phase protein, MBL-C plays an important role in the innate immune response. The aim of this study was to identify the key transcriptional factors in the promoter region (1 009 bp) of MBL2 gene, and explore the expression regulation mechanism of MBL2 gene. The promoter sequence 1 009 bp of MBL2 gene was selected from Hainan black goat, and a serial of 5'-flanking deletion regions of the goat MBL2 promoter were amplified and subcloned into pGL3-Basic vector. The recombined plasmids were transfected into 293T cells and detected with the dual-luciferase activity detection system to screen the core promoter region of MBL2 gene. The transcription factor binding site of the core promoter region of Hainan black goat MBL2 gene were predicted by online bioinformatics software. The site mutation technology and dual-luciferase activity detection system were utilized to identify the transcription factor binding sites. The results showed that the core promoter region of Hainan black goat MBL2 gene located in the range of -304 to -45 bp of the transcription initiation site. Online software analysis indicated that this region existed three transcription factors binding sites, including RELA, NF-κB2 and MZF1. Dual-luciferase report analysis results showed that the deletion of the binding sites of RELA and NF-κB2 extremely significantly reduced the transcriptional activity of goat MBL2 gene (P < 0.01). These results indicated that RELA and NF-κB2 might positively activate transcriptional regulation of goat MBL2 gene. This study can provide a theoretical basis for further exploring the function of MBL2 gene in Hainan black goat.

Key words: Hainan black goat, MBL2 gene, transcriptional regulation, dual-luciferase, core promoter

CLC Number:

S827.2

WNAG Xuemei, ZHAI Zhe, CHEN Qiaoling, WU Yanru, WU Haotian, HUANG Huixian, LIU Zhiyong, LI Chongrui, MANCHU Riga, WANG Fengyang, DU Li, CHEN Si. Screening and Analysis of Transcriptional Regulatory Elements of MBL2 Gene in Hainan Black Goat[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1795-1806.

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Screening and Analysis of Transcriptional Regulatory Elements of MBL2 Gene in Hainan Black Goat

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