Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (4): 304-312.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1224

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Gene Cloning, Expression Pattern, and Promoter Activity Analysis of Chicken BMP15

YANG Lan(), ZHANG Chen-xi, FAN Xue-wei, WANG Yang-guang, WANG Chun-xiu, LI Wen-ting()   

  1. College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450000
  • Received:2022-10-08 Online:2023-04-26 Published:2023-05-16

Abstract:

The purpose of this study is to investigate the transcriptional regulation mechanism of the bone morphogenetic protein 15(BMP15)gene in chickens, as well as its function, structure, and promoter active region. RACE(rapid amplification of cDNA ends)was used to clone the full-length cDNA of the chicken BMP15gene, and bioinformatics was to analyze the properties and structures of its encoding protein. Quantitative real-time PCR was plied to detect the expressions of chicken BMP15 gene in different tissues. Dual luciferase was used to identify the primary promoter region of the gene. According to the results, the chicken BMP15 gene has a transcript of 1 865 bp in length, encoding 350 amino acids, and its hereditary correlation is the closest to that of turkey. The protein structure primarily consists of disordered coils, there is a signal peptide region and no transmembrane domain. Additionally, BMP15 gene was significantly expressed in both ovaries and granulosa cells of the chicken(P<0.01)according to the constructed tissue-expression profile. Dual-luciferase assay revealed that the core promoter region of chicken BMP15 gene was positioned at -1 bp to -153 bp. This sets the foundations for further research into the BMP15 gene's function and its transcriptional regulatory mechanism.

Key words: Taihang chicken, BMP15 gene, bioinformatics, promoter activity