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PMID: 6316281 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis.

Nucleic acids research ·Vol. 11 ·No. 22 ·1983-11-25 ·Pages 8019-30

Nilsson B, Uhlén M, Josephson S, Gatenbeck S, Philipson L

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.

MeSH Terms
Ampicillin/toxicity Bacteriophage lambda/genetics Base Sequence DNA Transposable Elements Escherichia coli/genetics Genes, Bacterial Genes, Viral Genetic Vectors Mutation Oligodeoxyribonucleotides/chemical synthesis,genetics Oligonucleotides/genetics Penicillin Resistance Plasmids Transcription, Genetic
Chemicals
DNA Transposable Elements Oligodeoxyribonucleotides Oligonucleotides Ampicillin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Nilsson B
Uhlén M
Josephson S
Gatenbeck S
Philipson L
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23 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/11.22.8019
Published
1983-11-25
Pages
8019-30
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326556
Subset
IM
Databases
GENBANK
X00166
External Links
PubMed source DOI: 10.1093/nar/11.22.8019 PMC Full Text View journal details
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