抗菌肽CM4基因克隆及其在毕赤酵母中的表达鉴定  

Cloning, expression and characterization of antibacterial peptide CM4 in Pichia pastoris

作　　者：张杰[1] 吴希[1] 岳园园[1] 陈玉清[1] 张双全[1] 

机构地区：[1]南京师范大学生命科学学院江苏省分子医学生物技术重点实验室,南京210097

出　　处：《微生物学报》2005年第5期720-723,共4页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30271093)~~

摘　　要：为了在毕赤酵母中获得抗菌肽CM4表达,将抗菌肽CM4基因克隆入表达质粒pPIC9,SacⅠ线性化的重组表达质粒pPIC9-CM4电击转化P.pastorisGS115(his-),采用MM、MD板和PCR法筛选Mut+表型,甲醇诱导表达;抗菌活性检测、Tricine-SDS-PAGE及酸性活性电泳均表明抗菌肽CM4在毕赤酵母中成功地分泌表达;重组抗菌肽CM4对黑曲霉(Aspergillus niger)、绿色木霉(Trichoderma viride)都有很强的抑菌活性。To explore a new approach of expression of ABP-CM4 in methylotrophic yeast P. pastoris. The gene of ABP- CM4 was cloned into the vector pPIC9. The Sac Ⅰ -linearized plasmid pPIC9-CM4 was transformed into P. pastoris GSll5 by electroporation. By means of MM and MD plates and PCR, the recombinant P.pastoris strains （his^- mut^＋ ） were obtained. Antibacterial assay, Tricine-SDS-PAGE and Polyarylamide Gel Electrophoresis demonstrated that ABP- CM4 was extracellularly expressed in P. pastoris with the induction of methanol. The recombinant ABP-CM4 has the antifungal activity on Aspergillus niger and Trichoderma viride.

关 键 词：抗菌肽CM4 毕赤酵母 表达 pPIC9 真菌 

分 类 号：Q789[生物学—分子生物学]