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Volume 146, Issue 5

Isolation of a novel insertion sequence from using a trap vector based on inactivation of a reporter gene

The EMBL accession number for the sequence reported in this paper is Y18875 MF018875

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Abstract

An insertion sequence of has been isolated using a trap vector following insertion in and inactivation of the reporter gene. The trap vector is a temperature-sensitive () –mycobacterium shuttle plasmid, pCD4, which contains M, the kanamycin-resistance gene as a selection marker and a expression cassette. The mutation present in pCD4 functions in mycobacteria and enables screening for transposable elements from the mycobacterial genome that disrupt the gene by screening for white colonies on X-Gal plates in both mycobacterial as well as hosts. The vector was used to isolate a novel 1653 kb insertion sequence from named IS. IS duplicated host DNA at the target site, had inverted repeats at its ends and contained two ORFs on one strand. One of the predicted proteins showed homology to a putative transposase from . IS was present in two copies in the genome of . The trap vector appears to be useful in trapping insertion sequences from different mycobacteria by screening for the disrupted LacZ phenotype.

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  • Accepted:
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  • Published Online:
Keyword(s): insertion sequence , IS, insertion sequence , Kan, kanamycin; ts, temperature sensitive , molecular trap , Mycobacterium fortuitum and reporter gene
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2000-05-01
2024-04-19
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Isolation of a novel insertion sequence from Mycobacterium fortuitum using a trap vector based on inactivation of a lacZ reporter geneThe EMBL accession number for the sequence reported in this paper is Y18875MF018875
Microbiology 146, 1157 (2000); https://doi.org/10.1099/00221287-146-5-1157
/content/journal/micro/10.1099/00221287-146-5-1157
/content/journal/micro/10.1099/00221287-146-5-1157
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