| 摘要 |
PURPOSE:To obtain a complex plasmid vector pHY244PLK capable of carrying out multiple copy in each of Escherichia coli and Bacillus subtitis by cutting a complex plasmid pHY253PLK with SnaB1, subjecting the cut complex plasmid to partial digest treatment with an exonuclease and then cyclizing the treated plasmid. CONSTITUTION:Plasmid pHY300PLK is subjected to partial cutting treatment with AccI and then cutting treatment with BanI and XboI linker [d(pC-C-T-C-G- A-G-G)] is inserted therein to cyclize these ingredients and provide a complex plasmid vector pHY250PLK. Two pair of bases are site-specifically inserted into a controlled domain of replicated gene repalpha1 of the above-mentioned complex plasmid vector in Bacillus subtitis to form SnaB1 recognition cutting site and provide a complex plasmid vector pHY253PLK. The treatment mentioned in the above-mentioned purpose sentence is applied to the complex plasmid vector pHY253PLK to provide the complex plasmid vector pHY244PLK characterized by the restriction enzyme cutting map shown in the right figure. |
