Kuznedelov K D, Kolocheva T I, Rivkin M I, Kumarev V P
Bioorg Khim. 1986 Jun;12(6):842-4.
For subcloning separate synthetic gene fragments, a plasmid vector pSSC1 was constructed by inserting a synthetic polylinker into plasmid pBR 327 at the EcoRI-PstI sites. There are two FokI and HgaI sites at the ends of this polylinker in the opposite orientation, with the EcoRI and PstI sites between them. DNA fragments cloned at the EcoRI and PstI sites can be regenerated by either FokI or HgaI, the EcoRI and PstI sites being deleted from the cloned sequences. Such fragments have unique cohesive ends that allows their directed ligation into longer DNA (genes).
为了亚克隆单独的合成基因片段,通过将合成多克隆位点插入质粒pBR 327的EcoRI - PstI位点构建了质粒载体pSSC1。该多克隆位点两端有两个方向相反的FokI和HgaI位点,它们之间是EcoRI和PstI位点。在EcoRI和PstI位点克隆的DNA片段可以通过FokI或HgaI再生,克隆序列中的EcoRI和PstI位点会被删除。这些片段具有独特的粘性末端,能够将它们定向连接到更长的DNA(基因)中。