大连工业大学学报2025，Vol.44Issue(5)：325-331,7.DOI:10.19670/j.cnki.dlgydxxb.2025.0503
Paraburkholderia caffeinilytica CF1基因敲除体系的构建及转化条件优化
Construction of gene knockout system and optimization of transformation conditions of Paraburkholderia caffeinilytica CF1
摘要
Abstract
To establish the Paraburkholderia caffeinilytica CF1 knockout system,the helper plasmids were introduced into the wild-type strains based on the Red/ET homologous recombination technology.The knockout box with tetracycline resistance gene was constructed by overlap extension PCR,and the target gene was knocked out by introducing it into the wild-type strains.The positive knockout strains were obtained by multiple rounds of screening on the tetracycline resistance plate and sequenced for verification.The single factor test was used to optimize the conditions of gene knockout of strain CF1,such as the assisted plasmid,the preparation time of competent cells,the type of electroporation buffer,the concentration of recombinant fragments and the recovery time of the knockout strain,to achieve the efficient knockout of the target gene in strain CF1.The knockout system established in this experiment can also be used to knock out the target genes of other Paraburkholderia strains,providing an efficient genetic operation system for studying the gene function of Paraburkholderia strains.关键词
Paraburkholderia caffeinilytica/咖啡因/基因敲除体系/同源重组系统Key words
Paraburkholderia caffeinilytica/caffeine/gene knockout system/homologous recombi-nation system分类
生物科学引用本文复制引用
刘思佳,高子晴,李宪臻..Paraburkholderia caffeinilytica CF1基因敲除体系的构建及转化条件优化[J].大连工业大学学报,2025,44(5):325-331,7.基金项目
辽宁省教育厅高等学校基本科研项目(LJKMZ20220879). （LJKMZ20220879）
