摘要
旨在对沼泽型水牛CYP19A1基因进行克隆、生物信息学分析及对其在水牛组织的表达规律进行系统的探索。根据GenBank已公布的牛CYP19A1基因序列,设计特异性引物应用RT-PCR方法扩增克隆获得目的基因片段;应用生物信息学方法,分析和预测了水牛CYP19A1的理化性质、蛋白质二级及三级结构;应用QRT-PCR技术对CYP19A1在水牛组织的表达进行了差异分析。测序结果表明:水牛CYP19A1基因的编码区全长1 512bp,共编码503个氨基酸。多重序列比较分析显示水牛CYP19A1核苷酸序列与牛、绵羊、猪、人、马相应序列相似性分别为99%、98%、91%、86%和84%,结合系统进化树分析结果推测,CYP19A1基因在不同物种以及进化的过程中具有高度保守性。定量分析结果显示:水牛CYP19A1在水牛垂体、大脑、肝脏、骨骼、卵巢、肌肉和心脏组织具有不同程度的表达,其中垂体和大脑表达量最高,骨骼和卵巢次之,肌肉表达最低。本研究成功地克隆了沼泽型水牛CYP19A1基因并研究了其在不同水牛组织中的表达规律,为阐明其在水牛性腺中参与的激素合成转化规律等研究奠定了理论基础。
In order to clone, sequence bioinformatic analysis and expression pattern determination of CYP19A1 gene in swamp buffalo, a pair of special primers were designed according to released sequence of bovine CYP19A1 in GenBank. The CYP19A1 gene was amplified by RT-PCR, its gene sequence character and protein structure was systemically analysised by bioinformatics tech- niques. The expression patterns of buffalo CYP19A1 in different tissues were also assayed with qRT-PCR. The results showed that the cloned 1 587 bp buffalo CYP19A1 gene fragment inclu- ding a 1 512 bp whole length CDS （coded 503 amino acids）. The sequence multialigned results showed that buffalo CYP19A1 gene shared 99 ~, 98 %, 91~, 86 G and 84 ~ of similar nueleotide sequence with that of Bos taurus, Ovis aries, Sus scro f a and Homo sapiens, respectively. In ad- dition, the expression pattern analysis results showed that buffalo CYP19A1 expressed in all the seven detected tissues, with the most abundant expression in brain and pituitary gland, followed by skeleton and ovary, the minimal expression in muscle was observed. The cloning and analysis of CYP19A1 gene provided an important foundation for further study regulation mechanism of CYP19A1 gene in buffalo in the future.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2013年第4期514-521,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家转基因重大专项(2011ZX08007-003)
国家高技术研究发展计划(863)项目(2011AA100607)
国家自然基金项目(31160457)
