Showing papers by "Richard A. Jorgensen published in 1979"
TL;DR: A cleavage site map of Tn5 for restriction enzymes BamHI, B glI, BglII, Hind II, HindIII, HpaI, SalI, AvaI, SmAI, XhoI, PstI, PvuII, HaeII and HaeIII that was determined by the analysis of restriction enzyme cleavage patterns of ColEl.
Abstract: This paper reports a cleavage site map of Tn5 for restriction enzymes BamHI, Bg/I, Bg/II, Hind II, HindIII, HpaI, Sa/I, Aval, SmaI, XhoI, PstI, PvuII, HaeII and HaeIII that was determined by the analysis of restriction enzyme cleavage patterns of ColEl, two independent ColEl::Tn5 plasmids, and a ColEl::Tn5 deletion derivative. Ba/I, EcoRI, KpnI, and PvuI do not cleave Tn5. Construction and analysis of in vitro-generated deletions of a ColEl::Tn5 plasmid limit the sequences encoding neomycin resistance to a 1500-base-pair-long segment of Tn5. Insertion of DNA at a Bg/II site within this segment results in loss of the neomycin resistance phenotype. Since this Bg/II site lies in an inverted repeat region, sequences within this repeat seem to be involved in the expression of neomycin resistance.
531 citations
TL;DR: Results indicate the orientation of the tet gene and support the conclusion that the tet polypeptie is required for tetracycline resistance.
Abstract: The location of Tn10 genes encoding tetracycline resistance and its regulation was determined by analyzing the properties of recombinant plasmids carrying partial HpaI digestion products of lambda::Tn10 transducing phage deoxyribonucleic acid Within a 2,700-base pair region are encoded tetracycline resistance, the structural gene (tet) for a tetracycline-inducible polypeptide, and the regulatory elements for the induction of both the resistance phenotype and the polypeptide Fusion of different sequences to an HpaI site in the tet gene alters the molecular weight and stability of the polypeptide as well as the tetracycline resistance phenotype of strains producing fusion polypeptides These results indicate the orientation of the tet gene and support the conclusion that the tet polypeptie is required for tetracycline resistance A HincII cleavage site immediately upstream from the tet gene is protected by ribonucleic acid polymerase, but only the absence of ribonucleotide triphosphates The possibility that tet transcription is initiated at this site is discussed
117 citations
TL;DR: A cleavage map of a recombinant plasmid carrying Tn10 was constructed for 13 different restriction enzymes; it confirmed the previously reported structure of this transposon, which in part codes for the tetracycline resistance functions and is bounded by inverted repeats.
Abstract: A cleavage map of a recombinant plasmid carrying Tn10 was constructed for 13 different restriction enzymes. The Tn10 region of this plasmid contains cleavage sites for BamHI, AvaI, BglI, BglII, EcoRI, XbaI, HincII, HindIII, and HpaI. Restriction enzymes PstI, SmaI, KpnI, XhoI, SalI, and PvuI do not cleave within the Tn10 element. This map confirms the previously reported structure of this transposon; it is composed of a unique sequence (approximately6,400 base pairs long), which in part codes for the tetracycline resistance functions and is bounded by inverted repeats (approximately 1,450 base pairs long).
75 citations