摘要
利用根癌农杆菌LBA4404介导,建立了丝状真菌简青霉(Penicillium simpli-cissimum)H5的遗传转化系统.潮霉素抗性筛选、PCR和Southern blot分子鉴定等结果表明:筛选获得的转化子能够稳定遗传,外源的T-DNA以单拷贝随机整合到简青霉的基因组中.实验初步研究了农杆菌浓度、乙酰丁香酮浓度和共培养时间等因素对转化效率的影响,经过优化后转化体系的效率可达50个转化子/105个孢子.农杆菌介导的遗传转化方法在简青霉上的运用,将为研究该菌的基因工程改造提供强有力的工具.
Agrobacterium tumefaciens-mediated transformation（ATMT）was applied in Penicillium simplicissimum H5 by Agrobacterium tumefaciens LBA4404.The results of hygromycin-resistance selection,PCR and Southern blot analysis showed that T-DNA was randomly inserted into the transformants with single copy and the transformants indicated genetic stability.The transformation efficiency was about 50 transformants per 105 spores by optimizing the co-culture time,the concentration of Agrobacterium tumefacien and acetoyringone（AS）.The application of ATMT technique in Penicillium simplicissimum H5 will provide a powerful tool for the genetic engineering of this fungus.
出处
《湖南大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2010年第10期72-76,共5页
Journal of Hunan University:Natural Sciences
基金
留学回国人员科研启动基金资助项目(教外司留(2007)108号)
湖南省科学技术厅科技计划资助项目(2009WK2010)
关键词
根癌农杆菌
简青霉
转化
agrobacterium tumefaciens
penicillium simplicissimum
transformation
