| 摘要 |
NEW MATERIAL:A plasmid pNI 10 having about 3.7Kb molecular weight and enzyme cut map expressed by the formula and derived from the genus Pseudomonas which is not cloven by restriction enzymes BamHI, SacI and Sal I. USE:A vector for gene recombinant. PREPARATION:For example, Pseudomonas flavide IF-4 (FERM P-10865) strain is inoculated into a cultue medium and subjected to shake culture one night at 30 deg.C and the bacteria are collected by cooling and centrifuging the cultured liquid and suspended in a buffer and lysozyme, pronase and RNAase are added thereto and allowed to stand on ice and cooled and centrifuged to separate supernatant and isopropanol is added to supernatant and the resultant precipitate is separated to give crude DNA specimen and ethidium bromide and cesium chloride are added to the crude DNA specimen and the mixture is subjected to super centrifugation to separate a plasmid DNA, which is then subjected to electrophoresis and part having same moving degree as straight-chain DNA of 2.3Kb is dissolved and plasmid DNA is absorbed into a filter by sucking and filtering the solution with a filter to provide the pNI 10 plasmid. |
