Mettus A M, Macaluso A
Ecogen Inc., Langhorne, Pennsylvania 19047-1810.
Appl Environ Microbiol. 1990 Apr;56(4):1128-34. doi: 10.1128/aem.56.4.1128-1134.1990.
Bacillus thuringiensis delta-endotoxin (crystal protein) genes are normally expressed only during sporulation. It is possible to produce crystal protein during vegetative growth by placing B. thuringiensis crystal protein genes downstream of a strong vegetative promoter. By removing a possible transcriptional terminator of the tetracycline resistance gene of pBC16 and inserting a multiple cloning site, delta-endotoxin genes can be cloned downstream from the tetracycline resistance gene promoter. This construct allows for readthrough transcription from the strong vegetative promoter. Crystal protein is then produced during vegetative growth as well as during sporulation in both B. thuringiensis and Bacillus megaterium. This construct also allows for production of delta-endotoxin in B. thuringiensis strains that do not normally produce delta-endotoxin because of a defect in sporulation.
苏云金芽孢杆菌δ-内毒素(晶体蛋白)基因通常仅在孢子形成过程中表达。通过将苏云金芽孢杆菌晶体蛋白基因置于强营养启动子的下游,有可能在营养生长期间产生晶体蛋白。通过去除pBC16的四环素抗性基因的一个可能的转录终止子并插入一个多克隆位点,δ-内毒素基因可以克隆到四环素抗性基因启动子的下游。这种构建体允许从强营养启动子进行通读转录。然后,在苏云金芽孢杆菌和巨大芽孢杆菌的营养生长期间以及孢子形成期间都会产生晶体蛋白。这种构建体还允许在由于孢子形成缺陷而通常不产生δ-内毒素的苏云金芽孢杆菌菌株中产生δ-内毒素。