Efficient transposition of Tn 4556 by alterations in inverted repeats using a delivery vector carrying a counter-selectable marker for Streptomyces

Author:

Sota Masahiro1,Sakoda Akiko1,Ikeda Haruo2

Affiliation:

1. grid.484496.3 NAGASE R&D CENTER, NAGASE & CO., LTD. 651-2241 Kobe Hyogo Japan

2. 0000 0000 9206 2938 grid.410786.c Kitasato Institute for Life Sciences Kitasato University 252-0373 Sagamihara Kanagawa Japan

Abstract

Abstract A 6625-base pair transposon, Tn 4556, 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 Tn 4556 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 Tn 4556 transposition was markedly facilitated using a delivery vector carrying a strictly counter-selectable marker for Streptomyces strains.

Publisher

Oxford University Press (OUP)

Subject

Applied Microbiology and Biotechnology,Biotechnology,Bioengineering

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