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- Title
鸡gga-miR-31-5p 启动子真核表达载体的构建及其转录因子结合位点预测.
- Authors
邓哲宇; 王乙婷; 王颖洁; 胡菜; 吴宇慧; 赵宗仪; 左其生; 张亚妮
- 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.
- Subjects
PROMOTERS (Genetics); TRANSCRIPTION factors; BINDING sites; CELL lines; GENE transfection; PLASMIDS
- Publication
Acta Agriculturae Zhejiangensis, 2022, Vol 34, Issue 4, p713
- ISSN
1004-1524
- Publication type
Article
- DOI
10.3969/j.issn.1004-1524.2022.04.07