摘要
载体pTG402与地衣芽孢杆菌噬菌体B1p7经限制酶酶切并连接后转化大肠杆菌。从全部转化子中抽提质粒DNA,转化枯草杆菌后经菌落原位显色选到22个有启动子功能片段的克隆子。利用邻苯二酚双加氧酶对底物的显色反应,测定了15个克隆子的启动子活性,并绘制了启动子功能最强的重组质粒的酶切图谱。此外,还测定了两个克隆子在枯草杆菌各个生长期的表达情况,发现在对数生长后期表达量大增,认为识别这两个启动子的σ因子可能是σ^(37)。
Phage Blp7 DNA was digested with restriction endonuclease andligated to the restriction endonuclease digested tector pTG402. The ligatedwas used to transform comrelent cells of E.coli NC1061. Plasmid DNAminture was extracted from pooled transformants and competent cells ofB.subtilis were transformed. By selecting yellow colonies upon sprayingwith catechol solution,22 clones containing DNA fragments with promoterfunction were obtained. The promoter activity of 15 clones were deter-mined by the color reaction of catechol 2,3-dioxygenase. The insertedfragnent of the most potent promoter was mapped with restrictionenzymes. CatO_2ase activity of two clones were measured in cells of B. subtilis of all growth phases and was found to increase rapidly at theend of the log phase. It is inferred that these two promoters might berecognized by sigma 37.
出处
《生物工程学报》
CAS
CSCD
北大核心
1991年第1期11-17,共7页
Chinese Journal of Biotechnology
关键词
启动子
地衣芽孢杆菌
噬菌体
克隆
Promoter
temperate phage of Bacillus licheniformis
catechol 2, 3-dioxygenase
sigma factor
