RRC ID 33494
Author Nilsson B, Uhlén M, Josephson S, Gatenbeck S, Philipson L.
Title An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis.
Journal Nucleic Acids Res
Abstract 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.
Volume 11(22)
Pages 8019-30
Published 1983-11-25
DOI 10.1093/nar/11.22.8019
PMID 6316281
PMC PMC326556
MeSH Ampicillin / toxicity Bacteriophage lambda / genetics Base Sequence DNA Transposable Elements Escherichia coli / genetics* Genes, Bacterial Genes, Viral Genetic Vectors* Mutation* Oligodeoxyribonucleotides / chemical synthesis Oligodeoxyribonucleotides / genetics* Oligonucleotides / genetics* Penicillin Resistance Plasmids* Transcription, Genetic
IF 11.502
Times Cited 147
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Prokaryotes E. coli pUN122