An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis.
AUTOR(ES)
Nilsson, B
RESUMO
An Escherichia coli plasmid vector, pUN121, has been constructed which allows for positive selection of transformants harboring DNA inserts. The positive selection of transformants harboring DNA inserts. The vector is based on plasmid pTR262 (Roberts et al. Gene, 12, (1980), 123-127) in which the tetracycline resistance gene is under transcriptional control of the repressor protein coded by the phage lambda cI gene. This plasmid has been rearranged, using in vitro recombinant techniques including oligonucleotide mediated mutagenesis to yield a smaller plasmid (4.4 kb) with unique cloning sites for EcoRI, XmaI and SmaI in addition to the unique HindIII and BclI sites. The plasmid has a functional ampicillin resistance gene and the new restriction sites (EcoRI, XmaI and SmaI) when used for cloning, give rise to tetracycline resistant transformants.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=326556Documentos Relacionados
- High-efficiency oligonucleotide-directed plasmid mutagenesis.
- A simple vector modification to facilitate oligonucleotide-directed mutagenesis.
- Improved method for PCR-mediated site-directed mutagenesis.
- p21-ras effector domain mutants constructed by "cassette" mutagenesis.
- An improved thermal cycle for two-step PCR-based targeted mutagenesis.