鸡BMP15基因克隆、表达模式及启动子活性分析

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

为了解鸡骨形态生成蛋白15(bone morphogenetic protein 15,BMP15)基因的功能、结构及启动子活性区域,探究该基因的转录调控机制。通过RACE(rapid amplification of cDNA ends)技术克隆鸡BMP15基因cDNA全长序列,生物信息学分析其编码蛋白的性质及结构。实时荧光定量PCR(RT-qPCR)检测鸡BMP15基因在不同组织中的表达情况,双荧光素酶报告系统确定该基因的核心启动子区。结果表明,鸡BMP15基因存在一个长度为1865 bp的转录本,编码350个氨基酸,其遗传距离与火鸡最近。该蛋白为水溶性蛋白,其结构主要由无规则卷曲构成;存在信号肽区域,无跨膜结构域。构建的组织表达谱结果表明,BMP15基因在鸡的卵巢组织和颗粒细胞中表达极显著(P<0.01)。双荧光报告结果显示,BMP15基因启动子-153--1 bp为核心区域。为进一步确定BMP15基因功能及探究其转录调控机制奠定基础。

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 BMP15 gene,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 1865 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.