Construction of new vectors for cloning of promoters

Vectors for cloning promoter-DNA fragments were derived from plasmid pBR313 (Bolivar et al., 1977). These have several unique restriction sites and carry the trpA gene from Escherichia coli as a selective marker. The selection is based on an enhancement of the growth rate of those bacteria in which...

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Bibliographic Details
Published inGene Vol. 15; no. 4; pp. 297 - 305
Main Authors Enger-Valk, B.E., van Rotterdam, J., Kos, A., Pouwels, P.H.
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.01.1981
Subjects
Cloning, Molecular
DNA Restriction Enzymes - metabolism
DNA, Bacterial
Escherichia coli - genetics
Genetic Markers
Genetic Vectors
Operon
Plasmids
Substrate Specificity
Tryptophan - genetics
tryptophan gene
Plasmids
bp
kb
tryptophan gene
TSase A
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Summary:Vectors for cloning promoter-DNA fragments were derived from plasmid pBR313 (Bolivar et al., 1977). These have several unique restriction sites and carry the trpA gene from Escherichia coli as a selective marker. The selection is based on an enhancement of the growth rate of those bacteria in which the expression of trpA is directed by the cloned promoter. The expression of trpA can be determined quantitatively, independently of the copy number of the vector, and should reflect the apparent strength of the promoter, since the DNA segment located before trpA contains translational stop signals in all three reading frames.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
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ISSN:0378-1119
1879-0038
DOI:10.1016/0378-1119(81)90173-6