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Target sequence specificity of transposon Tn5 in the absence of major hotspots in the plasmid pBR322: identification of a new hotspot

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1006/plas.1993.1043
AuthorSearch for: ; Search for: 1; Search for: 1
Affiliation
  1. National Research Council of Canada. NRC Plant Biotechnology Institute
FormatText, Article
Abstract
The plasmid pLB11-1 is a pBR322 derivative in which part of the tetracycline resistance (tet) gene (basepair coordinates 23 to 375), containing five hotspots for Tn5 insertion (D. E. Berg et al., Genetics105, 813-828, 1983), has been replaced with a 5.1-kb fragment of Escherichia coli genomic DNA encoding an osmoregulatory function. Restriction mapping of 40 pLB11-1::Tn5 derivatives, chosen at random from 240 Tn5 insertion derivatives with an unaltered osmotolerant phenotype, placed Tn5 in the vector portion of 20 clones. The majority of these insertions (16/20) were located in a 0.09-kb region immediately downstream of the β-lactamase (bla) gene. Nucleotide (nt) sequence analysis of seven insertions from this group revealed an identical site of insertion within pBR322, representing a hitherto unidentified hotspot for Tn₅ insertion. The target sequence, 5′-GTCTGACGC, was found to be duplicated in these cases.
Publication date
PublisherElsevier
In
  • Plasmid 30, no. 2 (September 1993): 155–158.
LanguageEnglish
Peer reviewedYes
IdentifierS0147619X83710437
NRCC-36487
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Record identifier1b828d7a-91ce-458e-9476-84565ccf7829
Record created2023-11-06
Record modified2023-11-06
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