Construction of eukaryotic expression vector of chicken gga-miR-31-5p promoter and prediction of its transcription factor binding sites

doi:10.3969/j.issn.1004-1524.2022.04.07

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (4): 713-719.DOI: 10.3969/j.issn.1004-1524.2022.04.07

• Animal Science • Previous Articles Next Articles

Construction of eukaryotic expression vector of chicken gga-miR-31-5p promoter and prediction of its transcription factor binding sites

DENG Zheyu(), WANG Yiting, WANG Yingjie, HU Cai, WU Yuhui, ZHAO Zongyi, ZUO Qisheng, ZHANG Yani()

Key Laboratory of Animal Genetics Breeding and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, Jiangsu,China

Received:2020-10-12 Online:2022-04-25 Published:2022-04-28
Contact: ZHANG Yani

Abstract

Abstract:

The purpose of this study was to determine the basic active region of chicken gga-miR-31-5p promoter, analyze the important transcription factors regulating its transcription and explore the expression and regulation mechanism of chicken gga-miR-31-5p. The promoter sequence of chicken gga-miR-31-5p was amplified by PCR and inserted into the pEGFP-N1 vector to construct the recombinant vector. The positive recombinant plasmids were identified by PCR with bacterial solution and sequencing, and then transfected into DF-1 cell lines to observe their priming activity. The transcription factor binding sites in gga-miR-31-5p promoter region were predicted using online prediction software. The results showed that the gga-miR-31-5p promoter fragment was cloned and its pEGFP recombinant vector was constructed successfully. The transfection results in vitro showed that the region had promoter activity, and important transcription factor binding sites such as RARα, ER, c-Jun and GR were found in the promoter region of chicken gga-miR-31-5p. The above results indicated that the promoter region of chicken gga-miR-31-5p was preliminarily determined, and the important transcription factor binding sites were found in this region, which lays a foundation for further study on the biological function of gga-miR-31-5p promoter.

Key words: chicken, gga-miR-31-5p, promoter, transcription factors

CLC Number:

S831

DENG Zheyu, WANG Yiting, WANG Yingjie, HU Cai, WU Yuhui, ZHAO Zongyi, ZUO Qisheng, ZHANG Yani. Construction of eukaryotic expression vector of chicken gga-miR-31-5p promoter and prediction of its transcription factor binding sites[J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 713-719.

Figures/Tables 5

References 19

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Construction of eukaryotic expression vector of chicken gga-miR-31-5p promoter and prediction of its transcription factor binding sites

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