Hepc20在毕赤酵母中的胞内表达及鉴定  

Intracellular expression and identification of Hepc 20 in Pichia pastoris

作　　者：Farzana RASHID 袁其朋[1] 孙新晓[1] 李飞[1] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029

出　　处：《北京化工大学学报（自然科学版）》2010年第2期104-108,共5页Journal of Beijing University of Chemical Technology(Natural Science Edition)

基　　金：国家自然科学基金(20576010)

摘　　要：构建了Hepc 20的毕赤酵母表达载体,在毕赤酵母中能成功表达有活性的Hepc 20。优化了菌株的培养诱导条件,结果表明BMMY培养基是Hepc 20表达和重组菌株生长的最佳培养基,甲醇诱导终体积分数0.5%,在此条件下Hepc 20在重组菌株中的表达量约为3.6 mg/L。Western blot检测显示的22000处的条带为重组Hepc 20条带;ELISA验证表明重组Hepc 20可以跟抗体特异结合,琼脂扩散法抗菌实验表明重组Hepc 20对金黄色葡萄球菌和枯草芽孢杆菌表现出抗菌活性。A recombinant Pichia pastoris strain for Hepc 20 production has been constructed and an active hepcidin peptide was successfully expressed in methylotrophic yeast. A buffered methanol complex （BMMY） medium was found to be optimal for the hepcidin protein expression and growth of the recombinant strains, and when the final volume fraction of methanol was 0.5% , the expression level of hepcidin protein reached the maximum value of 3.6 mg/L. Western blotting showed the 22000 band of the recombinant hepcidin protein whilst enzyme-linked immu- nosorbent （ELISA） assay showed that the recombinant Hepc 20 peptide can combine with specific antibodies, and the agar diffusion technique showed that Hepc 20 exhibited antibacterial activity against S. aureus and B. subtilis.

关 键 词：抗菌肽 Hepc 20 铁 毕赤酵母 

分 类 号：Q786[生物学—分子生物学]