论文部分内容阅读
本研究旨在构建1个适用于苹果炭疽叶枯病菌(Glomerella cingulata)的遗传转化载体,为系统研究该病原菌基因的功能提供便利。通过基因工程及分子生物学技术,将三磷酸甘油醛脱氢酶(GAPDH)基因启动子Pgap和新霉素磷酸转移酶基因(nptⅡ)连结于质粒p Cambia0380中。构建的载体p Gapneo R1导入根癌农杆菌LBA4404后,通过农杆菌介导的转化技术(ATMT),成功将nptⅡ基因盒整合到苹果炭疽叶枯病菌基因组中。Southern blot分析结果表明,随机挑取的转化子T-DNA均以单拷贝插入到苹果炭疽叶枯病菌基因组中,表明该载体适合于苹果炭疽叶枯病菌的遗传转化。此外,该载体也可用于构建苹果炭疽叶枯病菌的目标基因的回补载体、超表达载体和荧光融合蛋白表达载体。
This study aimed to construct a genetic transformation vector suitable for Glomus cingulata to facilitate the systematic study of the function of the pathogen gene. The GAPDH promoter Pgap and the neptomycin phosphotransferase gene (nptII) were linked into plasmid pCambia0380 by genetic engineering and molecular biology techniques. After the vector p Gapneo R1 was introduced into Agrobacterium tumefaciens LBA4404, the nptII gene cassette was successfully integrated into the genome of apple bacterial blight by Agrobacterium-mediated transformation (ATMT). Southern blot analysis showed that T-DNA was randomly inserted into the genome of apple blight bacterial blight, indicating that the vector was suitable for the genetic transformation of apple blight. In addition, the vector can also be used to construct the gene recombination vector, the super-expression vector and the fluorescent fusion protein expression vector of the target gene of apple anthracnose blight bacterium.