大黄鱼与美国红鱼抗菌肽Piscidin串联基因的构建、酵母表达及抗菌活性鉴定  

Construction,yeast expression and antimicrobial activity identification of antimicrobial peptide Piscidintandom gene from Pseudosciaena crocea and Sciaenops ocellatus

作　　者：乔莹 王军[2] 马笑晚 潘滢 柯巧珍 郑炜强 苏永全[2] QIAO Ying;WANG Jun;MA Xiaowan;PAN Ying;KE Qiaozhen;ZHENG Weiqiang;SU Yongquan(Fourth Institute of Oceanography,Ministry of Natural Resources,Beihai 536000,China;College of Ocean and Earth Sciences,Xiamen University,Xiamen 361102,China;State Key Laboratory of Large Yellow Croaker Breeding,Ningde 352103,China)

机构地区：[1]自然资源部第四海洋研究所,广西北海536000 [2]厦门大学海洋与地球学院,福建厦门361102 [3]大黄鱼育种企业国家重点实验室,福建宁德352103

出　　处：《厦门大学学报（自然科学版）》2020年第1期43-48,I0002,I0003,共8页Journal of Xiamen University：Natural Science

基　　金：中央引导地方科技发展专项资金(2017L3019);大黄鱼技术平台建设项目(XDHT2018143A);国家海水鱼产业技术体系项目(CARS-47);自然资源部第四海洋研究所博士启动基金(201803);广西自然科学基金青年基金(2018JJB130206)

摘　　要：采用Piscidin基因串联表达的方案,通过Eco 31Ⅰ限制性内切酶定向连接方法合成Pseudosciaena crocea-Sciaenops ocellatus-piscidin(PSP)串联基因.以毕赤酵母(Pichia pastoris)的穿梭表达载体pPICZαA构建重组表达质粒pPICZαA-PSP,转化入毕赤酵母SMD1168菌株中.应用实时荧光定量PCR方法进行多拷贝克隆筛选,实现了PSP重组串联肽的诱导表达和纯化,并对其抑菌活性进行初步鉴定,为piscidin抗菌肽的生产应用奠定基础.In this study,the Pseudosciaena crocea-Sciaenops ocellatus-piscidin(PSP)tandom gene was constructed using a directional ligation method with Eco 31Ⅰrestriction endonuclease.The recombinant expression plasmid pPICZαA-PSP was constructed based on shuttle expression vector PPICZαA for Pichia pastoris,and then transformed into the P.pastoris strain SMD1168.The multi-copy transformants were screened using real-time fluorescent quantitative PCR,and the recombinant PSP tandom peptide was successfully induced expressed and purified,and its antimicrobial activity was preliminarily identified as well,providing basis for production and application of piscidin antimicrobial peptide.

关 键 词：抗菌肽 piscidin 串联基因 真核表达 抗菌活性 

分 类 号：Q781[生物学—分子生物学]