玉米圆斑病菌非核糖体肽合成酶6基因ATMT敲除载体构建  

Construction Deletion Vector of Nonribosomal Peptide Synthetases 6 of Helminthosporium carbonum

作　　者：张跃文[1] 张美丽[1] 张莉[1] 薛春生[1] 

机构地区：[1]沈阳农业大学植物保护学院,沈阳110161

出　　处：《玉米科学》2013年第4期127-131,共5页Journal of Maize Sciences

基　　金：教育部科学研究重点项目(211038);辽宁省自然科学基金(201102187)

摘　　要：以双元载体pPZP100为骨架,通过酶切、连接将标记基因(潮霉素-绿色荧光蛋白,HygR-GFP)及碳色长蠕孢(Helminthosporium carbonum)非核糖体肽合成酶基因(HcNPS6)侧翼序列插入目标载体pPZP100,构建HcNPS6基因敲除载体,并采用冻融法将pPZP100HygR-GFPHcNPS6转入农杆菌菌株AGL-1。挑取经氨苄/氯霉素筛选的重组子进行菌落PCR鉴定。结果表明,敲除载体已成功转入农杆菌AGL-1。Vector construction was based on the skeleton of pPZP100, HygR-GFP and flanking sequences of HcNPS6 of Helminthosporium carbonum were linked into pPZP100 by restriction enzymes digested and ligase connection. Deletion vector of nonribosomal peptide synthetases 6（NPS6） of Helminthosporium carbonum had been constructed pPZP100HygR-GFPHcNPS6 and transformed into AGL-1 by method of freeze-thaw.

关 键 词：玉米圆斑病菌 非核糖体肽合成酶6 敲除载体 农杆菌介导 

分 类 号：S513.035[农业科学—作物学] S435.131