磁性纳米颗粒介导的椭圆小球藻转化体系的建立  

Establishment of Chlorella ellipsoidea transformation system mediated by magnetic nanoparticles

作　　者：曹苏珊 薛静[2] 陈绪清[2] 王倩楠 张秀海[2] 安贤惠 CAO Su-shan;XUE Jing;CHEN Xu-qing;WANG Qian-nan;ZHANG Xiu-hai;AN Xian-hui(College of Marine Life and Fisheries,Jiangsu Ocean University/Jiangsu Key Laboratory of Marine Bioresources and Eco-environment,Lianyungang 222005,China;Beijing Agro-Biotechnology Research Center,Beijing 100097,China)

机构地区：[1]江苏海洋大学海洋生命与水产学院/江苏省海洋生物资源与生态环境重点实验室,江苏连云港222005 [2]北京农业生物技术研究中心,北京100097

出　　处：《江苏农业学报》2020年第2期306-311,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2017YFD0501005);北京市农林科学院科技创新能力建设专项(KJCX20170415)。

摘　　要：旨在利用磁性纳米颗粒作为载体构建小球藻的新型转化方法,由于纳米载体技术已经被成功地应用于动、植物的遗传转化中,其主要优势是相对于传统转化技术更加便捷、高效。本研究首先利用磁性纳米颗粒吸附含有增强型绿色荧光蛋白基因(EGFP)的质粒载体pYBA1132,形成纳米颗粒-基因复合物,通过磁场作用对椭圆小球藻原生质体进行转化,进而观察EGFP基因在椭圆小球藻中的瞬时表达情况。结果表明,对椭圆小球藻细胞进行酶解处理并形成半原生质体/原生质体后,磁性纳米颗粒可以介导外源EGFP基因转入小球藻中,并能够进行瞬时表达,产生绿色荧光。Using magnetic nanoparticles as carriers,a new transformation method of Chlorella ellipsoidea was constructed.Nanocarrier technology has been successfully applied to genetic transformation of animals and plants,which is more convenient and efficient than traditional transformation technologies.In this study,magnetic nanoparticles were used to absorb plasmid vector pYBA1132 containing enhanced green fluorescent protein gene(EGFP),and the nanoparticles-gene complexes were formed.In addition,the protoplasts of Chlorella ellipsoidea were transformed by magnetic force,and the transient expression of EGFP in Chlorella ellipsoidea was observed.After the formation of semi-protoplasts/protoplasts in Chlorella ellipsoidea cells by enzymolysis treatment,the magnetic nanoparticles mediated the transfer of exogenous EGFP gene into Chlorella ellipsoidea,and green fluorescence was detected after transient expression.

关 键 词：磁性纳米颗粒 增强型绿色荧光蛋白基因(EGFP) 转化 瞬时表达 

分 类 号：Q785[生物学—分子生物学]