摘要
以pCAMBIA1300-N载体为骨架,成功构建了以绿色荧光蛋白(gfp)为报告基因,潮霉素(hph)为抗性筛选标记的载体pKPG,并利用根癌农杆菌介导转化系统,成功获得了能表达绿色荧光蛋白的重组灰葡萄孢菌。通过PCR检测转化子的绿色荧光蛋白基因和潮霉素抗性表达框,观察菌丝和分生孢子的荧光表型,以及gfp基因的Southern杂交验证,结果表明:被测转化子基因组中均成功整合了目的基因片段。
Employing gfp as a reporter gene and hygromycin gene （hph） as a selection marker, the recombinant vector pKPG was constructed and transformed into fresh conidia of Botrytis cinerea via Agrobacterium tumefaciens. Transformants were identified by PCR analysis of gfp and hph cassette, green fluorescence observation with microscope and Southera hybridization. Results confirmed that target genes were successfully integrated into the genome of Botrytis cinerea.
出处
《遗传》
CAS
CSCD
北大核心
2008年第4期515-520,共6页
Hereditas（Beijing)
基金
浙江省重大科技攻关项目(编号:2004C12035)资助~~
关键词
灰葡萄孢菌
根癌农杆菌
绿色荧光蛋白
Botrytis cinerea
Agrobacterium tumefaciens
green fluorescent protein
