梅花鹿CYP26B1基因过表达载体的构建及进化树分析

发布时间：2018-03-30 20:13

  本文选题：CYPB　切入点：载体构建　出处：《中国兽医学报》2017年09期

【摘要】：CYP26B1是调节体内视黄酸(retinoic acid,RA)水平的一个关键酶,广泛存在于哺乳动物器官和组织中,对软骨细胞的分化具有重要的调节作用。本试验参照NCBI GenBank中已发表的牛CYP26B1基因序列,设计特异性引物。利用PCR方法扩增CYP26B1基因的编码区,并与pGEM-T载体连接,经EcoRⅠ和XhoⅠ双酶切后与pcDNA3.1载体融合得到CYP26B1过表达重组质粒。经测序鉴定后,转染鹿茸软骨细胞,利用实时荧光定量PCR方法检测CYP26B1mRNA表达变化。应用MEGA软件的Test Neighbor-Joining Tree法绘制CYP26B1基因系统进化树并进行比对分析。结果显示,重组质粒pcDNA3.1-CYP26B1转染鹿茸软骨细胞后,CYP26B1的表达量显著升高(P0.05)。进化树分析结果表明,梅花鹿CYP26B1基因与牛的基因同源性最高。本研究成功构建了梅花鹿CYP26B1过表达载体,为进一步研究CYP26B1在鹿茸生长及再生中的作用奠定基础。
[Abstract]:CYP26B1 is a key enzyme in regulating retinoic acid (retinoic acid) in vivo. It is widely found in mammalian organs and tissues and plays an important role in regulating the differentiation of chondrocytes. This study refers to the sequence of bovine CYP26B1 gene published in NCBI GenBank. Specific primers were designed. The coding region of CYP26B1 gene was amplified by PCR and ligated with pGEM-T vector. The recombinant plasmid of CYP26B1 overexpression was obtained by double enzyme digestion of EcoR 鈪，

本文编号：1687477