山东农业大学学报（自然科学版）2017，Vol.48Issue(4)：491-496,6.DOI:10.3969/j.issn.1000-2324.2017.04.003
辣椒疫霉菌效应因子RxLR23克隆与基因敲除体系建立
Cloning of RxLR23 and Establishment of Gene Knockout System of Phytophthora Capsici
摘要
Abstract
In the process of infection of Phytophthora capsici, pathogenic bacteria can secrete a large number of RxLR effect factors to help pathogens interfering with the disease resistance of host plants. For a long time, the lack of efficient techniques for directed mutagenesis and gene replacement has seriously hampered genetic study on pathogenicity of Phytophthora pathogenicity. In order to understand the function of RxLR effectors in Phytophthora capsici, using the Phytophthora capsici bacterial strain SD33DNA as template, we designed primers to clone the sequence of gene RxLR23,and it was used as the template to design sgRNA to construct pYF2.3G-ribo-sgRNA recombinant vector and to design the homologous recombinant donor vector pBluescript Ⅱ KS+recombinant vector. The CRISPR/Cas9 system was used to knockout gene RxLR23, after G418 resistance screening and PCR amplification verification, 4 knockout mutants of gene RxLR23 were obtained successfully.关键词
辣椒疫霉/效应因子/CRISPR/cas9/基因敲除Key words
Phytophthora capsici/effector/CRISPR/cas9/gene knockout分类
农业科技引用本文复制引用
姜海滨,李京,许海青,陈茹雪,张修国..辣椒疫霉菌效应因子RxLR23克隆与基因敲除体系建立[J].山东农业大学学报（自然科学版）,2017,48(4):491-496,6.基金项目
国家大宗蔬菜产业技术体系(CARS-25-03B) （CARS-25-03B）
