Structure and function of plasmid ColK.
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Base Composition↗
Biomedical subjects
W Tacon
Publications and source records attributed to W Tacon.
The construction and characterisation of plasmid vectors suitable for the expression of all DNA phases under the control of the E. coli tryptophan promoter.
A DNA fragment flanked in the E. coli genome by Hinf I sites and containing the E. coli tryptophan promoter-operator and nucleotides specifying the leader sequence and first seven amino acids of trp E, has been cloned in the Hind III site of pBR322 with the aid of Hind III linkers. The Hind III site upstream from the trp promoter in the transcriptional sense was deleted to generate a Hind III cloning vector, designated pWT111, suitable for the expression of cloned DNA sequences as fusion products. Derivatives of this plasmid were constructed, designated pWT121 and pWT131, whose Hind III cloning sites differ with respect to their translation phasing relative to the initiator ATG of the trp E gene. Both pWT121 and pWT131 have a higher copy number per cell than pWT111. The tetracycline genes of all three plasmids are under trp promoter control and could be used to monitor the transcription of foreign genes inserted at the Hind III site of the vectors.
Cloning, Molecular↗
ColE plasmid replication in DNA polymerase I-deficient strains of Escherichia coli.
Replication of the non-conjugative plasmids ColE1, ColE2 and Col3 has been examined in a number of DNA polymerase I-deficient strains, two of which contain the amber mutation polA1 along with either of two temperature-sensitive supF amber suppressors. These latter two strains produce reduced amounts of DNA polymerase I polymerizing activity of similar, if not identical properties to that produced by polA+ strains. Our results indicate that the ColE plasmids require different amounts of DNA polymerase I for stable plasmid maintenance. Moreover whereas all three plasmids are maintained in a strain defective in the 5' leads to 3' exonuclease activity of DNA polymerase I, ColE2 and ColE3 are not stably maintained between 30 degrees and 43 degrees in a number of DNA POLYMERASE I-deficient strains that are temperature-sensitive for ColE1 replication.
Alleles↗