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

Sabrina Semprini, Cheryl C. Collins, Kaarin C. Goncz, Giuseppe Novelli, Dieter C. Gruenert

Research output: Contribution to journalArticlepeer-review

Abstract

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 pSABR 01 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.

Original languageEnglish
Pages (from-to)1275-1280
Number of pages6
JournalBiotechnology Letters
Volume25
Issue number15
DOIs
Publication statusPublished - Aug 2003

Keywords

  • Full-length cDNA cloning
  • Plasmid
  • Plasmid purification
  • pSABR 01
  • pUC19

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Microbiology
  • Bioengineering

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