| 摘要 |
<p>The invention comprises a method of assembling several DNA units in sequence in a DNA construct and all derivatives of this method. In particular the production of synthetic enzymes is contemplated. Each DNA unit is provided with the same restriction enzyme recognition site at its 5' and 3' ends. The restriction recognition site at its 3' end being combined with a recognition site for a DNA modification enzyme. A DNA construct having the same or a compatible accessible restriction site, as provided in the DNA unit, is cleaved at the restriction site by the appropriate restriction enzyme. The desired DNA unit is then inserted into the DNA construct, this ligated product subsequently being brought into contact with a DNA modification enzyme such that the restriction site at the 3' end of the inserted DNA unit is abolished. The ligated product is then cleaved at the remaining unmodified restriction recognition site and a subsequent DNA unit is inserted. This process is repeated introducing each desired DNA unit to give a DNA construct containing all the desired units in sequence.</p> |
