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抗菌肽MagaininⅡ的原核表达与抗菌活性分析
引用本文:卢艳敏,崔海信,王承民,何宏轩.抗菌肽MagaininⅡ的原核表达与抗菌活性分析[J].河北农业大学学报,2011,34(5):59-62,68.
作者姓名:卢艳敏  崔海信  王承民  何宏轩
作者单位:1. 中国农业科学院农业环境与可持续发展研究所,北京100081;衡水学院生命科学系,河北衡水053000
2. 中国农业科学院农业环境与可持续发展研究所,北京,100081
3. 中国科学院动物研究所动物生态与保护生物学院重点实验室,国家野生动物疫病研究中心,北京100101
基金项目:中国科学院绿色农业技术集成与发展中心创新方向项目
摘    要:根据MagaininⅡ氨基酸序列,选用大肠杆菌偏爱密码子,化学合成了2个DNA片段,利用重叠区互补PCR法得到MagaininⅡ的基因序列.构建了融合表达载体pET21 b-MagaininⅡ,转化大肠杆菌BL21(DE3)pLysS,IPTG诱导表达,表达产物经SDS-PAGE分析,在5.1 kD处可见目的蛋白条带,...

关 键 词:MagaininⅡ  抗菌肽  融合表达  抑菌活性

Prokaryotic expression and antimicrobial activity analysisi of Magainin Ⅱ gene
LU Yan-min,CUI Hai-xin,WANG Cheng-min,HE Hong-xuan.Prokaryotic expression and antimicrobial activity analysisi of Magainin Ⅱ gene[J].Journal of Agricultural University of Hebei,2011,34(5):59-62,68.
Authors:LU Yan-min  CUI Hai-xin  WANG Cheng-min  HE Hong-xuan
Institution:1.Institute of Environment and Sustainable Development on Agriculture,Chinese Academy of Agricultural Sciences,Beijing 100081,China;2.Department of Life Sciences,Hengshui University,Hengshui 053000,China;3.National Research Center for Wildlife Borne Diseases,Key Laboratory of Animal Ecology and Conservation Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing 100101,China)
Abstract:Two sequences were designed using codon preference of Escherichia coli and synthesized by chemical method according to amino acid sequence of MagaininⅡ.The whole MagaininⅡgene fragment was obtained by overlap extension PCR.The amplified fragment was inserted into pET-21b and the fusion expression vector pET21b-MagaininⅡ was constructed.The recombinant plasmid was transferred into E.coli BL21(DE3) plyS and induced by IPTG.SDS-PAGE and Western-blotting analysis showed that the recombinant had expressed a 5.1 kD specific protein in E.coli.The fusion protein was expressed mainly in a soluble form after induction with 0.4 mmol/L IPTG at 20 ℃for 12 h.The fusion protein was purified and then antibacterial activity was tested.The activity assay demonstrated that the antimicrobial peptide had antibacterial activities against E.coli and Staphylococcus aureus.
Keywords:MagaininⅡ  antibacterial peptide  fusion expression  antimicrobial activity
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