| 牛源IL-6基因克隆与生物信息学分析 |
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| 引用本文： | 吕亚南,刘静雨,姚竞杰,安志兴,白跃宇,张晓建.牛源IL-6基因克隆与生物信息学分析[J].中国畜牧兽医,2019,46(11):3163-3171. |
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| 作者姓名： | 吕亚南 刘静雨 姚竞杰 安志兴 白跃宇 张晓建 |
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| 作者单位： | 1. 河南科技学院动物科技学院, 新乡 453003;2. 河南省动物卫生监督所, 郑州 410100 |
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| 基金项目： | 河南省基础与前沿技术研究计划（152300410095）；NSFC-河南人才培养联合基金（U1504306）；2014年度高层次人才科研启动项目（206010615007） |
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| 摘 要： | 试验旨在克隆牛白细胞介素-6（IL-6）基因，并对其编码蛋白进行生物信息学分析，对荷斯坦奶牛进行尾根采血提取RNA后逆转录为cDNA，根据NCBI数据库中已知的牛IL-6基因mRNA序列设计特异性引物，应用RT-PCR技术克隆得到CDS区全长序列，利用生物信息学软件分析牛IL-6基因序列特征、同源性及编码产物的理化性质等。结果表明，试验成功克隆了牛IL-6基因CDS区，全长627 bp，分子质量为23.759 ku，共编码208个氨基酸，理论等电点为7.58，脂溶系数为93.37，属于亲水性蛋白；牛IL-6基因与猪、绵羊、大鼠、人、猫、犬、鸡、小鼠、马和兔的同源性分别为84.4%、96.2%、66.7%、77.5%、75.9%、79.3%、48.5%、67.0%、79.5%和67.5%；系统进化树分析发现，牛与绵羊亲缘关系最近，猪次之，鸡最远；在二级结构预测中，该蛋白无规则卷曲与α-螺旋分别为34.6%和60.1%；亚细胞定位于细胞质；该蛋白含有信号肽和跨膜螺旋结构。本研究为进一步分析牛IL-6基因的功能奠定了一定的理论基础。
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| 关 键 词： | 牛 IL-6基因 克隆 生物信息学分析 |
| 收稿时间： | 2019-07-01 |
Cloning and Bioinformatics Analysis of IL-6 Gene in Bovine |
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| LYU Yanan,LIU Jingyu,YAO Jingjie,AN Zhixing,BAI Yueyu,ZHANG Xiaojian.Cloning and Bioinformatics Analysis of IL-6 Gene in Bovine[J].China Animal Husbandry & Veterinary Medicine,2019,46(11):3163-3171. |
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| Authors: | LYU Yanan LIU Jingyu YAO Jingjie AN Zhixing BAI Yueyu ZHANG Xiaojian |
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| Institution: | 1. School of Animal Science, Henan Institute of Science and Technology, Xinxiang 453003, China;2. Animal Health Supervision of Henan Province, Zhengzhou 410100, China |
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| Abstract: | The aim of the experiment was to clone the bovine interleukin-6 (IL-6) gene and analyze its coding protein bioinformatically.RNA was extracted from the tail heel blood of Holstein cows and retrieved into the cDNA.Specific primers were designed according to the known bovine IL-6 gene sequence in NCBI database,the full-length sequence of CDS region was cloned by RT-PCR.The biological characteristics of bovine IL-6 gene sequence,homology and physicochemical properties of coding products were analyzed by bioinformatics software.The results showed that the bovine IL-6 gene CDS region was successfully cloned,the total length was 627 bp,and the molecular weight was 23.759 ku,encoding a total of 208 amino acids.The theoretical isoelectric point was 7.58,the fat soluble coefficient was 93.37,which belonged to the hydrophilic protein.The homologies of Bos taurus IL-6 gene with Sus scrofa,Ovis aries,Rattus norvegicus,Homo sapiens,Felis catus,Canis lupus familiaris,Gallus gallus,Mus musculus,Equus caballus and Oryctolagus cuniculus were 84.4%,96.2%,66.7%,77.5%,75.9%,79.3%,48.5%,67.0%,79.5% and 67.5%,respectively.Phylogenetic tree analysis revealed that cattle and sheep had the closest genetic relationship,followed by pigs,and chickens was farthest.In the secondary structure prediction,the random coil and alpha-helix of the protein were 34.6% and 60.1% respectively.Subcellular localization was located in cytoplasm;The protein contained signal peptide and transmembrane helix structure.This study laid a theoretical foundation for further analysis of bovine IL-6 gene function. |
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| Keywords: | bovine IL-6 gene cloning bioinformatics analysis |
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