Cloning and expression of the 3' portion of the T4 denV gene as a lacZ fusion gene

Robert (Stephen) Lloyd, M. L. Augustine

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Polyclonal antibodies have been raised against endonuclease V from the bacteriophage T4. This rabbit serum, from which endemic E. coli antibodies have been removed, reacts with a single protein from T4-infected E. coli with a molecular weight of 6,078 dalton. It was confirmed that these antibodies were directed against endonuclease V through the inhibition of the pyrimidine dimer specific nicking activity of endonuclease V in an vitro nicking assay. A phage λgt11 T4 dC DNA library was screened for phage which produced a β-galactosidase-endonuclease V fusion protein. Immunopositive clones were detected at a frequency of 0.25% of the plaques in the library. Restriction enzyme analyses of the DNA from 45 of these phage showed that all contained a 1.8 kb T4 EcoRI fragment which had been inserted within λgt11 in a single orientation. Western analysis of proteins which were produced from an induction of lysogens made from these phage reveals a single fusion protein band with a molecular weight slightly larger than native β-galactosidase.

Original languageEnglish (US)
Title of host publicationMutation Research
Pages89-100
Number of pages12
Volume165
Edition2
StatePublished - 1986
Externally publishedYes

Fingerprint

Lac Operon
Deoxyribonuclease (Pyrimidine Dimer)
Organism Cloning
Galactosidases
Bacteriophages
Genes
Antibodies
Proteins
Molecular Weight
Escherichia coli
Pyrimidine Dimers
Restriction Mapping
Bacteriophage T4
Gene Library
Libraries
Clone Cells
Rabbits
DNA
Serum

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Lloyd, R. S., & Augustine, M. L. (1986). Cloning and expression of the 3' portion of the T4 denV gene as a lacZ fusion gene. In Mutation Research (2 ed., Vol. 165, pp. 89-100)

Cloning and expression of the 3' portion of the T4 denV gene as a lacZ fusion gene. / Lloyd, Robert (Stephen); Augustine, M. L.

Mutation Research. Vol. 165 2. ed. 1986. p. 89-100.

Research output: Chapter in Book/Report/Conference proceedingChapter

Lloyd, RS & Augustine, ML 1986, Cloning and expression of the 3' portion of the T4 denV gene as a lacZ fusion gene. in Mutation Research. 2 edn, vol. 165, pp. 89-100.
Lloyd RS, Augustine ML. Cloning and expression of the 3' portion of the T4 denV gene as a lacZ fusion gene. In Mutation Research. 2 ed. Vol. 165. 1986. p. 89-100
Lloyd, Robert (Stephen) ; Augustine, M. L. / Cloning and expression of the 3' portion of the T4 denV gene as a lacZ fusion gene. Mutation Research. Vol. 165 2. ed. 1986. pp. 89-100
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