【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。更多还原

【Abstract】 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 SacⅠ-linearized plasmid pPIC9-CM4 was transformed into P. pastoris GS115 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.更多还原