| 摘要 |
PURPOSE:To provide a plasmid vector for promoter searching use, containing an alpha-amylase structural gene and an incision site near the upstream of the structural gene with a restriction enzyme having no incision site in the sequence of said gene. CONSTITUTION:An alpha-amylase gene fragment of 2.0kbp containing BamHI site and prepared from plasmid pTUB4 containing an alpha-amylase structural gene and pHY300PLK treated with BamHI are treated with a ligase to obtain a plasmid of 6.9kbp. The plasmid is digested with BamHI and linked again to obtain a plasmid pTUD102 in which one of the BamHI incision sites is depleted. Separately pTD107 is produced from SmaI-treated pHY300pLKDELTAE and a fragment of 2.0kbp prepared from pTUP106 containing SD sequence and an alpha-structure gene. A plasmid pTUD109 is prepared from SmaI-treated PHY300pLDDELTAE and a fragment of 2.0kbp originated from plasmid pTUD108 containing synthesized SD sequence. The obtained plasmid is used as the objective vector. |
