摘要
筛选获得促进大肠杆菌质粒扩增的最优氯霉素浓度。采用单因素控制变量法,通过质粒转化、摇菌、质粒提取、PCR扩增、琼脂糖凝胶电泳检测等方法,并设置菌液培养时间梯度与氯霉素浓度梯度,分析不同浓度的氯霉素对大肠杆菌质粒扩增的影响。结果显示未加氯霉素的大肠杆菌菌液培养18 h时,质粒浓度最高,为(605.425±63.241)ng/μL;菌液培养18 h时加入10~250μg/mL的氯霉素对质粒扩增皆具促进作用;且浓度为150μg/mL时,质粒的扩增效率最高,为(645.425±45.042)ng/μL。10~250μg/mL的氯霉素对大肠杆菌质粒扩增皆具促进作用,促进作用最佳浓度为150μg/mL。
In order to obtain the optimal concentration of chloramphenicol for promoting the amplification of plasmid in Escherichia coli,this study set the time and the concentration gradient of chloramphenicol to explore the effects of different concentrations of chloramphenicol on the amplification of plasmid.Using single-factor control variable method,this paper performed plasmid transformation,shaking,plasmid extraction,PCR amplification,and agarose gel electrophoresis detection.When E.coli solution was not provided with chloramphenicol,the plasmid concentration had the highest(605.425±63.241 ng/μL)at 18 h of cultivation.Adding 10~250μg/mL chloramphenicol to culture for 18 hours could promote the amplification of plasmids.When the added concentration of chloramphenicol was 150μg/mL,the amplification efficiency got the highest(645.425±45.042)ng/μL.Chloramphenicol at the concentrations of 10-250 g/mL can promote the amplification of plasmid in E.coli,and the optimal concentration is 150μg/mL.
作者
陈绪美
尹志勇
檀军
田莹
李卓
郭建军
CHEN Xumei;YIN Zhiyong;TAN Jun;TIAN Ying;LI Zhuo;GUO Jianjun((Institute of Entomology, Guizhou University/Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region, Guiyang, Guizhou 550025, China)DIRFagopyrum cymosumDIRZanthoxylum piperitum inermeplatyrhyncha domestica)
出处
《山地农业生物学报》
2020年第6期81-84,共4页
Journal of Mountain Agriculture and Biology
基金
资源昆虫学研究科技创新团队(黔教合人才团队字[2015]71)
贵州省昆虫信息系统与资源开发利用重点实验室(黔科合LH字[2015]7685号)
国家自然科学基金项目(81803968)。
关键词
氯霉素
大肠杆菌
质粒
扩增
浓度
chloramphenicol
Escherichia coli
plasmid
amplification
concentration
