Title

Effect of IS900 gene of Mycobacterium paratuberculosis on Mycobacterium smegmatis

Authors

Authors

S. A. Naser; R. F. Gillespie; N. A. Naser;F. A. K. El-Zaatari

Comments

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Abbreviated Journal Title

Curr. Microbiol.

Keywords

CROHNS-DISEASE; IRON TRANSPORT; SEQUENCE; Microbiology

Abstract

Mycobacterium paratuberculosis is mycobactin dependent and contains multiple copies of the IS900 gene that encodes for p43 (46.5K protein). The correlation between the two characteristics has been investigated. A3.2-kb BamHI fragment from M. paratuberculosis containing the 1.451 kb IS900 gene was cloned in Escherichia coli and Mycobacterium smegmatis with pcDNA II and pNEZ6.3 plasmids, respectively. Surprisingly, the recombinant M, smegmatis grew poorly and slower in 7H9 broth supplemented with OADC (12 day) compared with M. smegmatis wild type or to M. smegmatis transformed with pNEZ6.3 (2 day). The growth rate of the recombinant M. smegmatis was restored by the addition of 2.4 mu M ferric mycobactin J to the media. There was no effect on the growth rate of E. coli recombinants. Western blot analysis with p43-specific anti-peptide antibodies resulted in the expression of 46.5K and a cleaved form of 33.5K protein bands in the recombinant E. coli. There was no expression in the recombinant M. smegmatis. A lower expression of 33.5K protein band was detected in the native M. paratuberculosis protein. The nucleotide sequence of the 3.2-kb fragment confirmed the presence of p43-encoded ORE There was no additional encoding sequence in the fragment. This suggests that the IS900 gene and/or its encoding products are involved in mycobactin dependency and possibly the slow growth rate of M. paratuberculosis.

Journal Title

Current Microbiology

Volume

37

Issue/Number

6

Publication Date

1-1-1998

Document Type

Article

Language

English

First Page

373

Last Page

379

WOS Identifier

WOS:000077055800003

ISSN

0343-8651

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