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细胞穿透肽 CCL 融合蛋白的构建与表达

刘勇 安艳芳 王仲霞 杨建业 吴君

中国感染控制杂志2016,Vol.15Issue(6):361-366,6.
中国感染控制杂志2016,Vol.15Issue(6):361-366,6.DOI:10.3969/j.issn.1671-9638.2016.06.001

细胞穿透肽 CCL 融合蛋白的构建与表达

Construction and expression of cell-penetrating peptide CCL fusion protein expression vector

刘勇 1安艳芳 2王仲霞 3杨建业 3吴君3

作者信息

  • 1. 南方医科大学深圳医院,广东 深圳 518101
  • 2. 湖北医药学院附属人民医院,湖北 十堰 442000
  • 3. 湖北医药学院附属人民医院,湖北 十堰 442000
  • 折叠

摘要

Abstract

Objective To evaluate the construction of expression vector for fusion protein of cell-penetrating pep-tide CCL (PEP-CCL).Methods CCL6-PEP-6XHis was inserted into plasmid pABP,pABP-CCL6-PEP plasmid was extracted and then transfected into HEK293 cells,CCL6-PEP-6XHis was expressed and purified by chromatog-raphy and detected with Western Blot.Results PEP-CCL express vector was successfully constructed and purified. PCR product of CCL6-PEP-6XHis Tag was ligated with T vector,recombinant was transferred into the host cells, then host cells were cultured,plasmid was extracted and sequenced,the sequence was identical to targeted gene. CCL6-PEP-6XHis was successfully inserted into the eukaryotic expression vector pABP,plasmid was extracted and digested,electrophoresis results revealed that a fragment with 430bp was digested by Hind Ⅲ+XbaⅠ,which was identical to the expected value.Western Blot revealed that CCL6-PEP fusion protein could be recognized by His monoclonal antibody.Conclusion PEP-CCL express vector can be constructed and expressed in eukaryotic cells.

关键词

细胞穿透肽/CCL/基因表达/重组融合蛋白质

Key words

cell-penetrating peptide/CCL/gene expression/recombinant fusion protein

分类

医药卫生

引用本文复制引用

刘勇,安艳芳,王仲霞,杨建业,吴君..细胞穿透肽 CCL 融合蛋白的构建与表达[J].中国感染控制杂志,2016,15(6):361-366,6.

基金项目

湖北省教育厅重点项目支持 ()

中国感染控制杂志

OA北大核心CSTPCD

1671-9638

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