Transposition burst of the ISH27 insertion element family inHalobacterium halobium

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Abstract
Investigation of the plasmid pHH4 in single colonies of Halobacterium halobium PHH4 indicated transposition of insertion elements in 20%of the colonies. Seven ISH27 insertions were observed as well as one ISH23 insertion. The various copies of ISH27 were compared to the two ISH27 elements already present In pHH4, and to the ISH27 element that was identified in the bacterio-opsin (bop) gene of a Bop mutant. These ten copies of ISH27 constitute three types on the basis of DNA sequence identity: ISH27-1 (1398 bp), ISH27-2, and ISH27-3 (1389 bp each). The DNA sequence comparison between the three types indicates a region of 1200 bp where the identity between ISH27-1 and ISH27-2 or ISH27-3 is 82–83%. ISH27-2 and ISH27-3 are 95% identical in this region. The remaining region exhibits a lower DNA similarity (64–74% Identity) between the different copies. An open reading frame of 1167 nucleotldes spans the more conserved region, and a corresponding transcript could be detected in H. halobium PHH4, but not in H. halobium wild-type. ISH27-1 is 91% identical to members of the insertion sequence-like elements ISH51 of Haloferax volcanll, whereas the other two ISH27 element types are 82–83% identical to ISH51. The transposition ‘burst’ of ISH27 was only seen after storage of the cells for more than two years at 4°C. Upon continuous cultivation at 37°C no transposition event could be observed, suggesting that stress factor(s) might have caused the high transposition rate.