侧孢短芽孢杆菌S62-9抗菌肽基因的初步定位  

Genetic locus mapping of antimicrobial peptide gene in Brevibacillus laterosporus S62-9

作　　者：张楠[1] 刘洋[1] 柯晓静[1] 于宏伟[1] 郭润芳[1] 贾英民[1,2] 

机构地区：[1]河北农业大学食品科技学院,河北保定071000 [2]河北科技大学生物科学与工程学院,河北石家庄050018

出　　处：《河北农业大学学报》2016年第5期82-86,共5页Journal of Hebei Agricultural University

基　　金：国家科技支撑计划(2013BAD10B03);河北省教育厅项目(QN2015029);河北省研究生创新项目

摘　　要：侧孢短芽孢杆菌S62-9可以产生高效广谱抑菌活性的细菌肽,为了确定该抗菌肽基因的遗传定位,本研究采用质粒消除和质粒转化进行正反论证。结果发现,电击法消除S62-9菌株的野生质粒pBLS62后,其突变株抑菌活性消失,而且对S62-9细菌肽的敏感性大大增强;而野生质粒pBLS62导入没有抑菌活性的枯草芽孢杆菌Wb800中,转化子Z4产生了抑菌活性,而且RP-HPLC结果表明Z4表达的抗菌物质与原始菌株S62-9抗菌肽为同一产物。从质粒消除和质粒转化与由此带来的表观性状的关系上,可以推断,侧孢短芽孢杆菌S62-9细菌肽及其免疫相关基因位于质粒而不是染色体上。Brevibacillus laterosporus S62-9is responsible for producing an antimicrobial peptide with broad-spectrum antimicrobial activity.The aim of this study is to confirm the genetic locus of antimicrobial peptide gene in B.laterosporus S62-9by plasmid curing and plasmid transforming techniques.The wild plasmid pBLS62 of the S62-9strain was eliminated by electric stroke,which led to the loss of the antimicrobial activity of the plasmid cured derivatives against Staphylococcus aureus SA137 and becoming extremely susceptible to AMP.The wild plasmid pBLS62 was transformed into Bacillus subtilis Wb800 and made it display the antimicrobial activity against S.aureus SA137.Furthermore,the results from RP-HPLC demonstrated the expression production of Z4 was the same as the antimicrobial peptide produced by B.laterosporus S62-9.All these results suggested that the antimicrobial peptide synthesis-related genes of B.laterosporus S62-9locate in the plasmid rather than other parts of the chromosome.

关 键 词：侧孢短芽孢杆菌 细菌肽 质粒消除 质粒转化 遗传定位 

分 类 号：Q781[生物学—分子生物学]