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Scientific article
English

Plasmid vectors for selecting IS1-promoted deletions in cloned DNA: sequence analysis of the omega interposon

Published inGene, vol. 103, no. 1, p. 17-23
Publication date1991
Abstract

We have constructed two plasmid vectors which allow selection for in vivo deletions within cloned DNA fragments. The plasmids are derivatives of pBR322 which carry the Escherichia coli rpsL (strA) gene, known to confer a dominant streptomycin (Sm)-sensitivity phenotype to the host cell, and a copy of the IS1 transposable element. Sm-resistant strains that harbor these plasmids display sensitivity to Sm. Spontaneous IS1-promoted deletions across the rpsL gene can be isolated simply by selection for Sm resistance. Hence, nested sets of deletions of a cloned DNA can be obtained and sequenced with an IS1-specific primer. Using this approach, we have determined the complete nucleotide sequence of the Ω interposon [Prentki and Krisch, Gene 29 (1984) 303–313].

Keywords
  • Streptomycin sensitivity
  • rpsL inactivation
  • Sequencing
  • Transposon
  • aadA
  • Recombinant DNA
Funding
  • Swiss National Science Foundation - 31-9073.87
  • Swiss National Science Foundation - 31-9438.88
Citation (ISO format)
PRENTKI, Pierre, BINDA, Anna, EPSTEIN, Andrée. Plasmid vectors for selecting IS<i>1</i>-promoted deletions in cloned DNA: sequence analysis of the <i>omega</i> interposon. In: Gene, 1991, vol. 103, n° 1, p. 17–23. doi: 10.1016/0378-1119(91)90385-O
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Article (Published version)
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ISSN of the journal0378-1119
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