| 摘要 |
PURPOSE:To provide a new insertion factor which is a transfer factor having a DNA base sequence originated from coryneform bacteria and utilizable for the preparation of insertion mutant, gene mapping, search for promoter, insertion of gene information, destruction of specific gene, etc. CONSTITUTION:Coryneform bacteria (e.g. Corynebacterium glutamicum ATCC 31831) is cultured in a medium to the prophase of logarithmic growth phase and the obtained bacterial cells are collected, treated successively with lysozyme and protease K and subjected to bacteriolysis by the addition of a surfactant. Total DNA is recovered from the lysed solution by conventional method. The DNA is amplified by PCR reaction using a primer consisting of a part of the base sequence of sacB and sacR gene regions of Bacillus subtilis and treated with restriction enzymes to obtain the objective new insertion sequence utilizable for the preparation of insertion mutant, gene mapping, search for promoter, insertion of gene information, destruction of specific gene, etc. |
