摘要
为了获得具有高抗菌活性和低溶血活性的抗菌肽,利用家蝇抗菌肽Cec Md和中国林蛙抗菌肽Chensirin,设计出杂合抗菌肽Cec Md-Che Rc,并构其克隆载体。选取Cec Md的N端和Chensirin的C端氨基酸序列组成杂合肽,通过互联网和计算机软件,分析比较杂合肽的特征参数并进行结构预测。采用重复序列PCR方法合成2条杂合肽Cec Md-Che Rc I和Cec Md-Che Rc IV的基因,并连接到克隆载体pMD18-T上。分析发现杂合抗菌肽中Cec Md-Che Rc I疏水性最强,Cec Md-Che Rc IV净正电荷数最高;两者N端为疏水性α螺旋结构,C端为亲水性α螺旋结构;两者疏水性氨基酸大体分布在螺旋轮的一侧,其余氨基酸分布在另一侧;带正电荷的氨基酸主要分布在亲水面上。2条杂合肽的基因大小分别为133 bp和124 bp;PCR、双酶切和测序鉴定表明成功地构建了克隆载体pMD18-T/Cec Md-Che Rc I和pMD18-T/Cec Md-Che Rc IV,为后续研究奠定了基础。
In order to acquire the new hybrid antibacterial peptide Cec Md-Che Rc with the more highly antibacterial activity and the lowliest erythrolysis activity, hybrid antibacterial peptide Cec Md-Che Rc were designed and got by the computer aided design, using Musca domestica antibacterial peptide and Rana cheminensis antibacterial peptide Chensirin, and the cloning vector of hybrid antibacterial peptide Cec Md-Che Rc were constructed. Hybrid antibacterial peptide Cec Md-Che Rc were made by Choose Amino acid sequences of Cec Md N terminal and chensirin C terminal. Amino acid sequences and characteristic parameters of hybrid antibacterial peptides were compared and analysised by the Internet and computer softwares. Two hybrid antibacterial peptides Cec Md-Che Rc I and Cec Md-Che Rc IV gene were synthesized using repetitive-element PCR and then inserted into cloning vector pMDI8-T.The results showed that there was the most highly hydrophobicity in the Cec Md-Che Rc I and the most positive charge was in the Cec Md-Che Rc IV; Both peptides have the hydrophobic N terminal and the hydropilic C terminal and form a or-helix structure in their two terminals; The hydrophobic amino acids of Cec Md-Che Rc I and Cec Md-Che Rc IV were distributed in the one side of helix wheel and other amino acids in another side;there are the most positive charge amino acid in hydropilic face; The genes of two peptides have 133 bp and 124 bp,respectively; The results of PCR,double enzyme digestion and Sequencing showed that two recombinant plasmids were constructed successfully. Two recombinant plasmids were constructed and the hybrid antibacterial peptides will be used for the future study.
出处
《黑龙江八一农垦大学学报》
2009年第6期47-50,67,共5页
journal of heilongjiang bayi agricultural university
基金
黑龙江省农垦总局"十一五"科技攻关项目(HNKXIV-09-02-05)
黑龙江省研究生创新科研资金项目(YJCX2009-134HLJ)
黑龙江八一农垦大学博士启动基金(校启B2009-10)
关键词
抗菌肽
分子设计
克隆载体
antibacterial peptide
design
cloning vector
作者简介
孙艳发（1981～），男，黑龙江八一农垦大学2007级硕士研究生。
通讯作者：张爱忠，男，教授，硕士研究生导师，E-mail：aizhzhang@sina．com
