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

ASJC Scopus subject areas

  • Medicine(all)

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    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)