DNA sequence of IS91 and identification of the transposase gene

IS91 is a 1,830-bp insertion sequence that inserts specifically at the sequence CAAG or GAAC of the target and does not duplicate any sequence upon insertion (23). By transposon mutagenesis, we have identified open reading frame 426 (ORF426; bp 454 to 1731) as the putative ORF for the transposase. It displays a cysteine-rich, potential metal-binding domain in its N-terminal region. Adjacent to ORF426, there is an ORF (ORF121) which precedes and terminally overlaps ORF426 by one amino acid. Tn1732 insertions in ORF121 do not affect the transposition frequency. IS91 has sequence similarities to IS801 from Pseudomonas syringae. Their putative transposases are 36% identical, including conservation of the cysteine-rich cluster. The information concerning IS801 insertion specificity and target duplication has been reevaluated in the light of our results.

[1]  D. Hanahan Studies on transformation of Escherichia coli with plasmids. , 1983, Journal of molecular biology.

[2]  J. Devereux,et al.  A comprehensive set of sequence analysis programs for the VAX , 1984, Nucleic Acids Res..

[3]  J. Walker,et al.  Distantly related sequences in the alpha‐ and beta‐subunits of ATP synthase, myosin, kinases and other ATP‐requiring enzymes and a common nucleotide binding fold. , 1982, The EMBO journal.

[4]  N. Datta,et al.  Trimethoprim resistance conferred by W plasmids in Enterobacteriaceae. , 1972, Journal of general microbiology.

[5]  F. Cruz,et al.  Specificity of insertion of IS91, an insertion sequence present in α‐haemolysin plasmids of Escherichia coli , 1989, Molecular microbiology.

[6]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[7]  M. Romantschuk,et al.  IS801, an insertion sequence element isolated from Pseudomonas syringae pathovar phaseolicola , 1991, Molecular microbiology.

[8]  P. R. Sibbald,et al.  The P-loop--a common motif in ATP- and GTP-binding proteins. , 1990, Trends in biochemical sciences.

[9]  D. Galas Bacterial insertion sequences. , 1989 .

[10]  Paul J. Hagerman,et al.  Sequence-directed curvature of DNA , 1986, Nature.

[11]  J. Trowsdale,et al.  A novel gysteine-rich sequence motif , 1991, Cell.

[12]  F. de la Cruz,et al.  Transposition of IS91 does not generate a target duplication , 1987, Journal of bacteriology.

[13]  R. Miller,et al.  A rapid and convenient method for the preparation and storage of competent bacterial cells. , 1988, Nucleic acids research.

[14]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[15]  F. de la Cruz,et al.  pACYC184-derived cloning vectors containing the multiple cloning site and lacZ alpha reporter gene of pUC8/9 and pUC18/19 plasmids. , 1988, Gene.

[16]  O. Fayet,et al.  Transposition in Shigella dysenteriae: isolation and analysis of IS911, a new member of the IS3 group of insertion sequences , 1990, Journal of bacteriology.

[17]  I. Dodd,et al.  Improved detection of helix-turn-helix DNA-binding motifs in protein sequences. , 1990, Nucleic acids research.

[18]  F. de la Cruz,et al.  Genetic and molecular characterization of Tn21, a multiple resistance transposon from R100.1 , 1982, Journal of bacteriology.

[19]  P Argos,et al.  The integrase family of site‐specific recombinases: regional similarities and global diversity. , 1986, The EMBO journal.

[20]  M. Kröger,et al.  IS150: distribution, nucleotide sequence and phylogenetic relationships of a new E. coli insertion element. , 1988, Nucleic acids research.

[21]  O. Fayet,et al.  Functional similarities between retroviruses and the IS3 family of bacterial insertion sequences? , 1990, Molecular microbiology.

[22]  E N Trifonov,et al.  A computer algorithm for testing potential prokaryotic terminators. , 1984, Nucleic acids research.

[23]  D. Lipman,et al.  Improved tools for biological sequence comparison. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. McKnight,et al.  The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. , 1988, Science.

[25]  F. de la Cruz,et al.  Several copies of the same insertion sequence are present in alpha-hemolytic plasmids belonging to four different incompatibility groups , 1982, Journal of bacteriology.

[26]  J. Berg,et al.  Zinc fingers and other metal-binding domains. Elements for interactions between macromolecules. , 1990, The Journal of biological chemistry.

[27]  R. Korneluk,et al.  Rapid and reliable dideoxy sequencing of double-stranded DNA. , 1985, Gene.

[28]  U. Rüther pUR 250 allows rapid chemical sequencing of both DNA strands of its inserts. , 1982, Nucleic acids research.

[29]  R. Schmitt,et al.  Tn1721 derivatives for transposon mutagenesis, restriction mapping and nucleotide sequence analysis. , 1986, Gene.