| 摘要 |
NEW MATERIAL:The plasmid pSAN181 having a molecular weight of 5 0.2 M-dalton, and incised by restriction enzymes BamHI, PstI, XhoIand BglII at single site each, wherein the incision sites with reference to the incision site by BamHIare 0, 0.85, 1.15 and 3.95 M-dalton, respectively. USE:Since the plamid has small molecular weight and single incision site for the restriction enzyme, it is useful as a vector for cloning. PROCESS:Steptomyces fulvoviridis SANK61781 strain (FERM-P No.6279) is cultured in a liquid medium under aerobic condition (preferably at 6-8pH and 25- 30 deg.C for 24-48hr), and mycelia are recovered from the product. The mycelia are suspended in a buffer solution containing EDTA, and subjected to bacteriolysis with cytolytic enzyme. A sodium chloride solution is added thereto, and the produced precipitate is removed. The resultant supernatant liquid is digested with ribonuclease and pronase to obtain DNA. The cyclic plasmid pSAN181 can be separated from the chromosome DNA. |
