Efficient transposition of Tn4556 by alterations in inverted repeats using a delivery vector carrying a
Masahiro Sota1, Akiko Sakoda1, Haruo Ikeda2
1NAGASE R&D CENTER, NAGASE & CO., LTD., Kobe, Hyogo, 651-2241, Japan.
Abstract:
A 6625-base pair transposon, Tn4556, was initially isolated from a Streptomyces strain and a sequence analysis was performed; however, its annotation data remain incomplete. At least three positions were identified as frameshift and base-exchange errors by resequencing. The revised sequence revealed that Tn4556 contains four open reading frames that encode transposase, methyltransferase, isoprenyl diphosphate transferase, and resolvase, respectively. Thirty-eight-base pair inverted repeat (IR) sequences at both ends contained a 1-bp mismatch flanked by a target duplication site, and transposition efficiency was improved by the replacement of imperfectly matched IR-L to perfectly matched IR-L. The detection of Tn4556 transposition was markedly facilitated using a delivery vector carrying a strictly counter-selectable marker for Streptomyces strains.
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