[1]颜欣欣,杭建雄,刘云秋,等.锡兰钩虫TIMP基因的序列分析与原核表达[J].中国预防兽医学报,2019,(09):969-972.[doi:0.3969/j.issn.1008-0589.201901004]
　YAN Xin-xin,HANG Jian-xiong,LIU Yun-qiu,et al.Sequence analysis and prokaryotic expression of Ancylostoma ceylanicum TIMP[J].Chinese journal of preventive veterinary medicine,2019,(09):969-972.[doi:0.3969/j.issn.1008-0589.201901004]
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锡兰钩虫TIMP基因的序列分析与原核表达()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年09
页码:
969-972
栏目:
研究简报
出版日期:
2019-10-25

文章信息/Info

Title:
Sequence analysis and prokaryotic expression of Ancylostoma ceylanicum TIMP
文章编号:
1008-0589(2019)09-0969-04
作者:
 

颜欣欣; 杭建雄; 刘云秋; 黄　岳; 冉荣坤; 赵　琪; 何　龙; 李　秀; 刘菊玫; 李国清*

 (华南农业大学 兽医学院，广东 广州 510642)
Author(s):
 

YAN Xin-xin;  HANG Jian-xiong;  LIU Yun-qiu;  HUANG Yue;  RAN Rong-kun;  ZHAO Qi; HE Long;  LI xiu;  LIU Ju-mei;  LI Guo-qing*

 (College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China)
关键词:
锡兰钩虫; 金属蛋白酶组织抑制剂; 基因克隆; 原核表达; 序列分析
Keywords:
Ancylostoma ceylanicum;  ;  tissue inhibitor of metalloproteinase (TIMP);  ;  cloning;  ;  prokaryotic expression;  phylogenetic analysis
分类号:
S852.7
DOI:
0.3969/j.issn.1008-0589.201901004
文献标志码:
A
摘要:
为表达锡兰钩虫金属蛋白酶组织抑制剂(Ace-TIMP)重组蛋白，本实验根据GenBank中锡兰钩虫TIMP (ANCCEY-04405)基因序列设计引物扩增犬源锡兰钩虫Ace-TIMP基因，利用生物信息学软件对其序列鉴定；根据编码的氨基酸序列进行遗传进化关系分析；构建其重组表达质粒pET-32a-Ace-TIMP-MP，转化大肠杆菌，并经诱导表达。结果显示，Ace-TIMP基因ORF为648 bp，编码215个氨基酸，包含16个氨基酸的信号肽，具有TIMP家族Cys-X-Cys特征性序列。进化树分析显示Ace-TIMP与犬钩虫TMP-2位于同一分支。经诱导表达与纯化，获得了分子质量约为42 ku的可溶性融合蛋白。本实验克隆了Ace-TIMP基因并利用原核表达系统表达了可溶性的重组蛋白，为Ace-TIMP生物学活性的研究和免疫潜能的评价奠定了基础。
　　
Abstract:
To prepare recombinant Ancylostoma ceylanicum tissue inhibitor of metalloproteinase (Ace-TIMP), primers were designed according to the sequence in GenBank (ANNECY-04405) to amplify the Ace-TIMP gene from dog-derived parasites. Bioinformatic softwares were used to identify the amplified sequence and phylogenetic analysis was conducted based on the deduced amino acid sequences. A plasmid for expressing Ace-TIMP (pET-32a-Ace-TIMP-MP) in E.coli was constructed to express recombinant protein. Results showed that the ORF of Ace-TIMP is 648bp and encodes 215 amino acids. Ace-TIMP contains a signal peptide with 16 amino acids and the characteristic sequence of TIMP family (Cys-X-Cys). Ace-TIMP is in the same branch with A.caninum TMP-2 in the phylogenetic tree. Finally, soluble fusion protein with molecular weight of about 42ku was expressed and purified from the induced E.coli. In this study, the coding sequence of Ace-TIMP was cloned and expressed through a prokaryotic expression system. The data laid a foundation for further study on biological activities of the protein and evaluation of its immunogenic potential.

参考文献/References:

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(本文编辑：李　爽；英文编辑：贾洪林)

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更新日期/Last Update: 2019-10-29