[1]刘春,陈晓虹,姜兰,等.鮰爱德华氏菌的红色荧光蛋白标记及其应用[J].江苏农业学报,2019,(03):661-666.[doi:doi:10.3969/j.issn.1000-4440.2019.03.022]
　LIU Chun,CHEN Xiao-hong,JIANG Lan,et al.Construction and application of Edwardsiella ictaluri strain labelled with mCherry fluorescent protein[J].,2019,(03):661-666.[doi:doi:10.3969/j.issn.1000-4440.2019.03.022]
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鮰爱德华氏菌的红色荧光蛋白标记及其应用()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年03期
页码:
661-666
栏目:
畜牧兽医·水产养殖
出版日期:
2019-06-30

文章信息/Info

Title:
Construction and application of Edwardsiella ictaluri strain labelled with mCherry fluorescent protein
作者:
刘春1; 陈晓虹1; 姜兰1; 曹际振1; 2; 李凯彬1; 王英英1; 常藕琴1; 王芳1; 石存斌1; 林明辉1; 王庆1
（1.中国水产科学研究院珠江水产研究所/农业部渔药创制重点实验室/广东省水产动物免疫技术重点实验室，广东广州510380；2.华南农业大学，广东广州510642）
Author(s):
LIU Chun1; CHEN Xiao-hong1; JIANG Lan1; CAO Ji-zhen1; 2; LI Kai-bin1; WANG Ying-ying1; CHANG Ou-qin1; WANG Fang1; SHI Cun-bin1; LIN Ming-hui1; WANG qing1
（1. Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences/ Key Laboratory of Fishery Drug Development of Ministry of Agriculture/Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Guangzhou 510380, China；2.South China Agricultural University, Guangzhou 510642, China）
关键词:
鮰爱德华氏菌; 红色荧光蛋白; 斑马鱼; 巨噬细胞
Keywords:
Edwardsiella ictaluri; mCherry fluorescent protein; zebrafish; macrophages
分类号:
S917.1
DOI:
doi:10.3969/j.issn.1000-4440.2019.03.022
文献标志码:
A
摘要:
为研究鮰爱德华氏菌与宿主的相互作用关系，本研究构建带有红色荧光蛋白基因的pMDmCherry表达载体，通过电击转化法将载体成功导入鮰爱德华氏菌zbl141菌株中，获得了红色荧光蛋白标记的鮰爱德华氏菌菌株。在倒置荧光显微镜下观察，重组鮰爱德华氏菌显示特异性红色荧光。稳定性检测结果表明，标记菌株传至28代，重组质粒稳定率仍为100%。用标记菌株感染斑马鱼仔鱼后，能在激光共聚焦显微镜下实时观察到细菌在鱼体内的分布情况；标记菌株体外感染小鼠巨噬细胞后，也能观察到病原菌与巨噬细胞的相互作用情况。说明标记菌株具有良好的特异性和可视性，为鮰爱德华氏菌对宿主的致病性研究提供了一种良好的生物材料。
Abstract:
To study the interaction between Edwardsiella ictaluri and its host, the pMDmCherry expression vector with mCherry fluorescent protein was constructed and transformed into E.ictaluri zbl141 strain by electroporation, and the recombinant E.ictaluri was obtained. The specific mCherry fluorescent could be observed in recombinant E.ictaluri under inverted fluorescence microscope. The results showed that the stability rate of recombinant plasmid was still 100% when the labelled strain was transferred to the 28th generation. The distribution of bacteria in zebrafish larva infected by the labelled strain could be observed in real time by confocal laser microscopy. After the mouse macrophages was infected by labelled strain in vitro, the interaction between pathogens and macrophages could be observed clearly. These results suggest that mCherrylabelled bacteria possess specificity and visibility, providing useful biological material for study of pathogenicity between E.ictaluri and the host.

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备注/Memo

备注/Memo:
收稿日期：2018-09-27 基金项目：中国水产科学研究院中央级公益性科研院所基本科研业务费专项（2017HY-ZC0406）；现代农业产业技术体系建设专项（CARS-46）；广东省科技计划项目(2016A030303028、2017A040403008) 作者简介：刘春（1982-），男，重庆九龙坡人，硕士，助理研究员，主要从事水产动物病害防治研究。（E-mail）lc5173483@163.com 通讯作者：王庆，（E-mail）sunny_929@163.com
更新日期/Last Update: 2019-06-30