| 摘要 |
PURPOSE:To provide a recombinant plasmid containing a human lysozyme gene having a DNA fragment at 5'-terminal, wherein said DNA fragment has a sequence to form a secondary structure which does not form a hydrogen bond between a translation initiation codon and a base complementing a ribosome-bonded site. CONSTITUTION:Oligodeoxyribonucleotides of formula I-formula IV are chemically synthesized and a DNA fragment of formula V is derived terefrom. A recombinant plasmid pPLHLY-1 containing human lysozyme gene is digested with a restruction enzyme TaqI, the obtained fragment containing human lysozyme gene is linked with the above DNA fragment and digested with Sau3AI and the obtained fragment is inserted into manifestation plasmid pMY12-6Amp-1 to construct pPLHLY-2. The product is cut with EcoRI and SalI and the obtained human lysozyme gene is integrated into the insertion site of plasmid vector pLY30, 32 to obtain a recombinant plasmid which can be copied in large number in a host cell, can be induced by temperature and gives a large amount of human lysozyme in high efficiency. |
