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A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance

✍ Scribed by Jorgensen, Richard A. ;Rothstein, Steven J. ;Reznikoff, Williams S.

Publisher: Springer
Year: 1979
Tongue: English
Weight: 657 KB
Volume: 177
Category: Article
ISSN: 0026-8925
DOI: 10.1007/bf00267254

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✦ Synopsis

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.

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