贵州黑山羊IFN-ε基因的克隆表达与生物信息学分析OA

[Objective]The structure and function of interferon-epsilon(IFN-ε)gene from Guizhou black goat were explored to provide the theoretical basis for prevention and control of goat infectious diseases and development of new antiviral drugs.[Method]Total RNA from the liver of Guizhou black goat was extracted,then the IFN-ε gene was amplified by RT-PCR and sequenced,and finally the gene sequence and encoded protein structure were analyzed and predicted by bioinformatics analysis.[Result]The length of the coding region of IFN-ε gene of Guizhou black goat was 582 bp,encoding 194 amino acids.The nucleotide sequence homology of Guizhou black goat IFN-ε gene with goat,sheep,takin and oryx was 99.7%,99.1%,99.1%and 97.9%,respectively.The amino acid sequence homology of Guizhou black goat IFN-ε gene with goat,sheep,takin and oryx was 99.0%,97.9%,97.9%and 94.8%separately.The double enzyme digestion result of the recombinant expression plasmid pET-28a-IFN-ε was correct.The size of the recombinant protein induced by the positive strain was approximately 39 ku.The recombinant protein was mainly expressed in a soluble protein form,and could specifically react with His-tagged mouse monoclonal antibody.The IFN-ε protein was an unstable hydrophilic protein with signal peptide and no transmembrane structure,and its subcellular localization was extracellular mainly.There were 16 phosphide acid sites and no glycosylation site in IFN-ε protein.The secondary structure of the IFN-ε protein was α-helix mainly.The IFN-ε protein mainly interacted with IL10RB,IFNAR2 and IFNAR1 proteins.[Conclusion]The sequence of the IFN-ε gene of Guizhou black goat is successfully cloned first.The structured recombinant expression plasmid pET-28a-IFN-ε is of a good immunogenicity.