| 摘要 |
PURPOSE:To provide the subject easily and safely available plasmid for controlling plant pathogenic bacteria, derived from a nonpathogenic mutant, M4S, of Pseudomonas solanacearum and capable of nonpathogenicizing Pseudomonas pathogenic bacteria. CONSTITUTION:M4S (FERM P-700), a nonpathogenic mutant of Pseudomonas solanacearum is inoculated into a medium followed by shaking culture at 30 deg.C for ca. 18hr and then subcultured. The resulting culture solution is centrifugated to collect microbial cells, which are, in turn, suspended in physiological saline followed by addition of a surfactant-contg. solution to effect bacteriolysis, and then plasmid is separated from the system by a conventional technique. The plasmid is then put to restriction enzyme treatment and incorporated with a drug-resistant gene and a promoter gene, thus obtaining the objective plasmid having 6.62kbp of molecular weight and broken sites due to EcoRI, StuI, XhoI, PstI, ApaI, HindIII or EcoRV, which can trans form pseudomonas pathogenic bacteria into nonpathogenic and nontoxic ones. |
