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Volume 146, Issue 9

ISBst, a novel type of insertion-sequence element causing loss of S-layer-gene expression in ATCC 12980

The GenBank accession number for the sequence reported in this paper is AF162268.

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Abstract

The cell surface of the surface layer (S-layer)-carrying strain of ATCC 12980 is completely covered with an oblique lattice composed of the S-layer protein SbsC. In the S-layer-deficient strain, theS-layer gene was still present but was interrupted by a novel type of insertion sequence (IS) element designated ISBst. The insertion site was found to be located within the coding region of the gene, 199 bp downstream from the translation start of SbsC. ISBst is 1612 bp long, bounded by 16 bp imperfect inverted repeats and flanked by a directly repeated 8 bp target sequence. ISBst contains an ORF of 1446 bp and is predicted to encode a putative transposase of 482 aa with a calculated theoretical molecular mass of 55562 Da and an isoelectric point of 913. The putative transposase does not exhibit a typical DDE motif but displays aHis-Arg-Tyr triad characteristic of the active site of integrases from the bacteriophage λ Int family. Furthermore, two overlapping leucine-zipper motifs were identified at the N-terminal part of the putative transposase. As revealed by Southern blotting, ISBst was present in multiple copies in the S-layer-deficient strain as well as in the S-layer-carrying strain. Northern blotting indicated that S-layer gene expression is already inhibited at the transcriptional level, since no -specific transcript could be identified in the S-layer-deficient strain. By using PCR, ISBst was also detected in PV72/p6, in its oxygen-induced strain variant PV72/p2 and in the S-layer-deficient strain PV72/T5.

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Keyword(s): Bacillus stearothermophilus , DIG-dUTP, digoxigenin-11-dUTP , insertion-sequence element , IS, insertion sequence , S-layer protein and S-layer, surface layer
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2000-09-01
2024-04-25
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ISBst12, a novel type of insertion-sequence element causing loss of S-layer-gene expression in Bacillus stearothermophilus ATCC 12980The GenBank accession number for the sequence reported in this paper is AF162268.
Microbiology 146, 2175 (2000); https://doi.org/10.1099/00221287-146-9-2175
/content/journal/micro/10.1099/00221287-146-9-2175
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