扬子鳄α型干扰素基因的克隆分析及表达量的测定  

Molecular cloning,sequence analysis and tissue expression detection of Chinese alligator IFN-α gene

作　　者：白艺兰 费荣梅[1] BAI Yi-lan;FEI Rong-mei(Key Laboratory of Animal Bacteriology,Ministry of Agriculture,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学农业部动物细菌开放实验室,江苏南京210095

出　　处：《中国兽医科学》2019年第4期461-467,共7页Chinese Veterinary Science

摘　　要：为研究扬子鳄IFN-α基因的序列、结构及生物学功能,对其进行了基因克隆、序列分析及表达量的测定。根据GenBank中登录的扬子鳄IFN-α基因序列(XM_006026085.1)设计合成特异性引物,通过RTPCR从成年扬子鳄外周血中扩增及克隆IFN-α基因CDS区,对其进行测序分析、进化树构建、同源性分析及结构和功能预测,并对扬子鳄不同组织中IFN-α的表达水平进行测定和分析。结果显示,扬子鳄IFN-α基因的完整ORF序列为672 bp,编码氨基酸223个,其中前26个氨基酸为信号肽。成熟蛋白的分子质量为26ku,等电点为9.51。三级结构预测显示该蛋白含有5段α螺旋,符合I型干扰素结构特征。同源性和进化树分析结果显示,扬子鳄IFN-α基因与密西西比鳄的同源性最高,在进化树中处于同一分支。IFN-α表达量在血液中最高,肌肉组织中表达量最低,非冬眠期的表达量高于冬眠期的表达量。上述研究结果为进一步研究扬子鳄IFN-α基因表达调控机制和基因功能奠定了基础。In order to study the sequence structure and biological functions of IFN-α gene,a pair of specific primers were designed to amplify the CDS region of Chinese alligator IFN-α gene.The sequence was analyzed by phylogenetic tree analysis,homology analysis and structural prediction.The differential expression of IFNp protein in different tissues were semiquantitatively analyzed during hibernation and non-hibernation.The resuIts showed that the complete ORF of Chinese alligator IFN-α gene was 672 bp,encoding 223 amino acids and the first 26 amino acids were signal peptides.In mature protei n,the molecular weight was 26 ku and the isoelectric point was 9.51.The 3D structure showed that the protein consisted of 5 alpha helixes.The IFNp gene of Chinese alligator was closest to that of Alligator mississippiensis.It was found that the highest expression level of IFN-α was in the blood and the lowest was in the muscle.The results laid the foundation for further studies on the mechanism of IFN-α gene expression regulation and functions of the gene.

关 键 词：IFN-Α 扬子鳄 序列分析 

分 类 号：S852.42[农业科学—基础兽医学]