芦花鸡IFITM3基因克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Interferon-induced Transmembrane Protein 3(IFITM3)Gene in Luhua Chicken

本研究旨在对芦花鸡的干扰素诱导的跨膜蛋白-3(interferon-induced transmembrane protein 3,IFITM3)基因进行克隆及生物信息学分析。根据GenBank中鸡IFITM3基因序列(登录号:NM_001350061.1)设计特异性引物,利用RT-PCR技术对芦花鸡IFITM3基因进行克隆、测序和生物信息学分析。结果表明,芦花鸡IFITM3基因片段大小为414bp,与GenBank中发表的鸡IFITM3基因序列(登录号:NM_001350061.1)同源性为99.9%;编码137个氨基酸。IFITM3基因理论分子质量为14.95ku,理论等电点(pI)为6.89,不稳定系数为33.98,脂肪系数为103.14,平均疏水指数为0.300,存在3个明显的疏水区,无信号肽,存在2个跨膜区,分别位于第46-68、101-123位氨基酸处;二级结构预测显示,IFITN3蛋白主要以无规则卷曲为主,存在多个α-螺旋和β-折叠区域。本研究成功克隆了芦花鸡IFITM3基因,为进一步研究IFITM3蛋白功能及阐明其抗病毒分子机制奠定了理论基础。

This study was aimed to clone the interferon-induced transmembrane protein 3（IFITM3）gene of Luhua chicken and analyze its genetic structure with bioinformatics.One pair of special primers were designed according to predicted sequence of chicken IFITM3 gene in GenBank（accession No.：NM-001350061.1）.The IFITM3 gene of Luhua chicken was amplified by RT-PCR,the physicochemical property and secondary structure of IFITM3 gene were systemically analyzed by bioinformatics.The results showed that the IFITM3 gene fragment included a414 bp whole length CDS（coding 137 bp amino acids）,and the gene sequence homology was above 99.9% comparison with the sequence published in GenBank（accession No.：NM-001350061.1）.The molecular weight,isoelectric point,instability index and fat coefficient of IFITM3 were 14.95 ku,6.89,33.98 and 103.14,respectively.The prediction result of IFITM3 gene structure showed that IFITM3 had no signal peptide but had three obvious hydrophobic domains,and the average hydrophobic index was 0.300.The two transmembrane domains might locate in the 46 to 68 and 101 to 123 amino acids across the online TMHMM 2.0 software.The prediction of secondary structure revealed that someα-helixes andβ-sheets exist in IFITM3 protein while random coil was major pattern.The results suggested that the whole CDS sequence of Luhua chicken IFITM3 gene was successfully cloned,which provided foundation for furtherstudying on the function and antiviral molecular mechanisms of Luhua chicken IFITM3 protein.