抗菌肽MagaininⅡ多拷贝串联体的设计与构建  

Design and Construction of Concatemer of Antibacterial Peptide MagaininⅡ

作　　者：成娟丽[1,2] 李锋[2] 王红[1] 林金水[3] 

机构地区：[1]运城学院生命科学系,山西运城044000 [2]武汉生物工程学院生物工程系,湖北武汉430415 [3]西北农林科技大学生命科学学院,陕西杨凌712100

出　　处：《生物技术》2014年第4期75-79,共5页Biotechnology

基　　金：国家青年科学基金项目("微生物来源的细菌烯脂酰ACP还原酶抑制剂的新型细胞筛选模型的建立与应用研究";No.31200040);校级博士科研启动项目("靶向细菌烯脂酰ACP还原酶的抗菌药物筛选模型的建立与应用";YQ-2011041)资助

摘　　要：目的:分析设计抗菌肽MagaininⅡ多拷贝串联体表达质粒并进行构建验证。方法:根据GeneBank中提供的抗菌肽MagaininⅡ氨基酸序列,结合表达菌株毕赤酵母密码子偏爱性,设计出MagaininⅡ的表达序列,并于其两端插入一对同尾酶SalⅠ和XhoⅠ的酶切识别序列及裂解位点AGA,人工合成后插入质粒载体pUC19中,利用同尾酶法构建抗菌肽MagaininⅡ多拷贝串联体。结果:经M13引物进行PCR及SalⅠ/PstⅠ双酶切进行鉴定,成功构建出抗菌肽MagaininⅡ1-5拷贝串联体重组质粒。结论:该研究设计的MagaininⅡ串联体构建方法可行。Objective： The antibacterial peptides Magainin Ⅱ gene was analysed and artificially constructed 5 copies of its concatemer. Method ： According to the amino acid sequence of antibacterial peptides Magainin H provided by GeneBank, this study has suc- cessfully designed the expression sequence of Magainin Ⅱ with a pair isocaudaners named Sal I ,Xho I and a cracking site AGA inserted in its terminal, combined with the anticodon preference of pichia. After synthesized artificially, the expression sequence is inerted to a plasmid carrier pUC19 to construct concatemer of antibacterial peptides Magainin Ⅱwith the use of isocandaners. Result ：5 copies concatemer of an- tibacterial peptides Magainin Ⅱ were confirmed by performing colony PCR utilizing primers M13 and the PCR products were digested with Sat I and Xho I . Conclusion ：The results that successfully constructed the antibacterial peptides Magainin Ⅱconeatemer had proved that the designing method was feasible.

关 键 词：抗菌肽 MagaininⅡ 同尾酶 串联体 

分 类 号：Q782[生物学—分子生物学]