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设计天蚕素抗菌肽在E.coli中的表达及活性鉴定

Expression Design Cecropin in E.Coli and Determine Antimicrobial Activity

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【作者】 韩晋辉翟培施用晖吕文平

【Author】 Han Jin-hui1,2,ZHAI Pei1,2,SHI Yong-hui1*,LU Wen-ping1(1.State Key Laboratory of Food Science and Technology,Ministry of Education,Jiangnan University,Wuxi 214122,China;2.Guangdong Food and Drug Vocational College,Guangzhou 510520,China)

【机构】 食品科学与技术国家重点实验室江南大学广东食品药品职业学院

【摘要】 为了研究天蚕素抗菌肽的构效关系,进一步提高其抗菌活性,收集并整理了62条天蚕素,通过生物信息学比对(blast),发现其中含有大量保守序列。在此保守序列的基础上,根据天蚕素结构特征,设计了4条抗菌肽。将设计抗菌肽的基因克隆到载体PUC19上,在大肠杆菌JM109中融合表达目的多肽,并促其形成包涵体。包涵体洗涤纯化后,用溴化氰裂解除去融合蛋白质,抑菌圈法测定其抗菌活性,结果表明其中抗菌肽AMP-CecC具有较强的抑菌活性。

【Abstract】 This manuscript was to investigate the relationship of structure and function of cecropins,for this,accumulated sixty-two cecropins,utilized bioinformatics blast cecropins were accumulated,and a large sum of consensus sequence was found.Base on these consensus sequences,the cecropins structural feature we design four new antimicrobial peptides to utilize technology of genetic engineering,gene of design antibacterial peptide cloned into vector PUC19,and expressed objective polypeptide in E.Coli JM109.We promote expression production to be inclusion.After cleaning inclusion,utilized cyanogen bromide(CNBr) we split fusion peptide and remove fusion polypeptide.Then detect its antimicrobial activity.The design antibacterial peptide AMP-CecC has strong antibacterial activity.

【基金】 中央级科研院所基础性工作专项资金重点项目(2001DEA20022)
  • 【文献出处】 食品与生物技术学报 ,Journal of Food Science and Biotechnology , 编辑部邮箱 ,2008年06期
  • 【分类号】Q78
  • 【被引频次】13
  • 【下载频次】314
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