摘要
对常规的农杆菌介导转化方法进行了改进,建立了一种高效构建耐受铜突变株的方法。与传统方法相比,该方法能够提高铜耐受菌株80%的筛选效率;同时,抗生素使用量减少80%。共获得15株高耐受铜的突变株,通过继代培养及纯化后,确定其耐受特性具有遗传稳定性。PCR与Southern blot验证了突变株确实为T-DNA随机插入。结果表明,该方法能够极大提高耐受铜突变株构建效率。
A modified Agrobacterium tumefaciens-mediated transformation method was established for the construction of mutants with improved copper tolerance and accumulation capability in Trichoderma reesei.According to the normal method,it can improve more than 80%efficiency.Furthermore,it can reduce about 80%antibiotics consumption.A total of 15 mutants were obtained by the modified method,which showed the genetic stability by subculture.PCR and Southern blot were detected the T-DNA insertion in the genome DNA of mutants.Results showed the modified method can improve obviously the construction efficiency of copper tolerant mutants.
作者
傅科鹤
范莉莉
李园园
熊国勇
FU Kehe;FAN Lili;LI Yuanyuan;XIONG Guoyong(Biology Department of Nanchang Normal University,330032,Nanchang,PRC)
出处
《江西科学》
2020年第2期179-182,225,共5页
Jiangxi Science
基金
国家自然科学基金地区项目(31660020)
江西省教育厅项目(151252,GJJ161233)。
关键词
木霉
耐铜
突变株构建
Trichoderma spp.
copper tolerance
mutants construction
