中国畜牧兽医

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布鲁氏菌外膜蛋白22基因克隆与原核表达

杜志强,林涛,刘晓燕,吴亚坤,王建英   

  1.  (内蒙古科技大学数理与生物工程学院,内蒙古包头  014010)

  • 收稿日期:2013-11-11 出版日期:2014-05-20 发布日期:2014-06-25
  • 通讯作者: 王建英(1945—),男,博士,教授,从事环境生物技术与布鲁氏菌病方面的研究。
  • 作者简介:杜志强(1980—),男,山西人,博士,副教授,研究方向:无脊椎动物先天免疫基因工程与布鲁氏菌病。
  • 基金资助:

    2011年内蒙古自治区自然科学基金(2011BS0502)。

Cloning and Prokaryotic Expression of Outer Membrane Protein 22 Gene in Brucella

DU Zhi-qiang, LIN Tao, LIU Xiao-yan, WU Ya-kun, WANG Jian-ying   

  1.  (School of Mathematics, Physics and Biological Engineering, Inner Mongolia University of Science and Technology, Baotou 014010, China)

  • Received:2013-11-11 Online:2014-05-20 Published:2014-06-25

摘要: 本试验以布鲁氏菌外膜蛋白22(omp22)基因作为研究对象,通过基因序列克隆、表达载体构建、大肠杆菌原核表达与亲和纯化,对蛋白的表达与纯化进行了研究。结果显示,omp22基因的核苷酸序列含有639 bp,编码212个氨基酸残基,预测分子质量为22 ku,电泳结果显示重组omp22蛋白的分子质量为47 ku,与理论值相符。本试验结果为进一步研究重组omp22蛋白的免疫刺激与免疫保护作用奠定了基础。

关键词:

布鲁氏菌; 外膜蛋白22基因; 基因克隆; 原核表达; 纯化

Abstract: In this experiment, the Brucella outer membrane protein 22 (omp22) gene was taken as the research object. Through the gene sequence cloning, expression vector construction, prokaryotic expression and affinity purification, expression and purification of the protein were studied. The results showed that the nucleotide sequence of omp22 gene contained 639 bp, which encoded 212 amino acids residues with predicted molecular weight of 22 ku. The electrophoresis results showed that molecular weight of recombinant omp22 protein was 47 ku, which was consistent with the theoretical value. All these results provided a good basis for immunity stimulation and immunity protective effect of recombinant omp22 protein in further research.

Key words:

Brucella; outer membrane protein 22 gene; gene cloning; prokaryotic expression; purification