克隆合作猪神经营养因子3基因(Neurotrophin3,NTF3)的编码区序列(Coding region sequence,CDS),预测分析其结构与功能,并检测基因在不同组织中的表达量,构建组织表达谱.通过PCR扩增、San-ger 测序等方法获得合作猪NTF3基因CDS区序列,使用生物信息学方法分析其结构,并使用RT-qPCR技术检测其在不同组织中的表达水平.结果显示,合作猪NTF3基因CDS区长774 bp,编码257个氨基酸;等电点(pI)为9.46,属于碱性蛋白;合作猪与猪、牛亲缘关系较近;二级结构主要为α-螺旋和无规则卷曲,三级结构主要由β-折叠、α-螺旋等组成;整条链中亲水性氨基酸残基数量多于疏水性氨基酸残基数量,是亲水性蛋白;无跨膜区域,是非跨膜蛋白;在37～58位氨基酸残基有一个线圈结构域、64～79位有一个低复杂度结构域和144～249位有一个神经生长因子结构域;存在信号肽,推测是分泌蛋白;NTF3基因在合作猪脾脏的表达量最高,回肠表达量最低.
Cloning,Bioinformatics Analysis and Tissue Expression Profile Construction of NTF3 Gene in Hezuo Pig
The coding region sequence(CDS)ofNTF3 gene in Hezuo pig was cloned to predict and an-alyze its structure and function,and expression levels in different tissues of Hezuo pig was detected to construct a tissue expression profile.The CDS region sequence of NTF3 gene in Hezuo pig was ob-tained by PCR amplification and Sanger sequencing,and analyzed by bioinformatics.The results showed that the CDS of NTF3 gene was 774 bp,encoding 257 amino acids.The isoelectric point(pI)was determined to be 9.46,belonging to alkaline protein.Hezuo pig had a close relationship with both pig and cattle.The secondary structure consisted of α-helix and random coil;while the tertiary struc-ture was mainly composed of β-sheet and α-helix.The number of hydrophilic amino acid residues in the whole chain was more than that of hydrophobic amino acid residues,indicating a predicted hydrophilic protein.There was no transmembrane region,non-transmembrane protein;there was a coiled coil re-gion domain between amino acid residues 37-58,a low complexity region domain at 64-79,and a nerve growth factor domain at 144-249,respectively.The NTF3 gene had the highest expression in the spleen and the lowest expression in the ileum of Hezuo pig.
万方数据
维普
