Construction and purification of pSABR 01, a pUC19-derived vector optimized for cloning full-length cDNA

摘要

A novel pUC19-derived vector, pSABR 01, was constructed by sub-cloning a fragment of the pSPORT1 polylinker into PUC19. The insertion of the polylinker generated two inactivating mutations in the LacZ open reading frame. These were then repaired by a PCR-based Site Directed Mutagenesis strategy. The pSABR01 plasmid has four sites that are recognized by 'rare-cutter' restriction endonucleases that will optimize the cloning of full-length cDNA and five dual restriction sites that increase the versatility of subcloning the inserted cDNA. Protocols were also defined for purification of pSABR 01 from residual pSPORT1, following pSABR 01 construction, and from another contaminating plasmid. [References: 9]