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Title of Journal: Curr Microbiol

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Abbravation: Current Microbiology

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Springer-Verlag

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DOI

10.1007/978-1-4613-1333-5_5

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1432-0991

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Efficient Transformation Procedure of a Newly Isol

Authors: Lotfi Mellouli Ines KarrayRebai Samiha Sioud Raoudha Ben AmeurMehdi Belgacem Naili Samir Bejar
Publish Date: 2004/12/31
Volume: 49, Issue: 6, Pages: 400-406
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Abstract

A new aerobic Grampositive bacterium designated TN58 producing antibacterial activities against Grampositive and Gramnegative bacteria was isolated from Tunisian soil The nucleotide sequence of the 16S rRNA gene 1516 bp of the TN58 strain showed high similarity 96–98 to the Streptomyces 16S rRNA genes especially with that of Streptomyces lavendulae which produces the antitumor compound mitomycin C and the cyclic peptide antibiotic complestatin Cultural characteristic studies alignment data of the 16S rRNA gene and analysis of the nucleotide sequence of a 22 kb genomic DNA fragment from TN58 strongly suggested that this strain could be an actinomycete and most probably belongs to the genus Streptomyces Study of the influence of different nutritional compounds on antibiotic production showed that the highest antibacterial activities were obtained when glycerol at 1 w/v was used as sole carbon source in the presence of potassium In analytical conditions the application to supernatant culture of the TN58 strain of various extraction and purification steps led to the isolation of two pure active molecules having a retention time of 386 and 502 min respectively TN58 strain was untransformable with the Streptomyces cloning vector pIJ702 via classical polyethylene glycol PEG protoplast transformation and previously described Streptomyces electroporation procedures Transformation was rendered possible by the electroporation technique only after utilization of a preculture medium without sucrose and a regeneration plate containing a low sucrose concentrationThis work was supported by the Tunisian government Contract Programme CBSLEMP We are grateful to Dr JL Pernodet BGM Laboratory of the IGM Orsay France for providing DNA probes from S ambofaciens and the M luteus LB 14110 strain and to Dr A Dhouib CBS for providing the four other indicator microorganisms Thanks are also due to Mr R Rekik Mr R Hmidi Mr E Ben Messaoud Dr M Ben Ali and Dr H Chouayekh for their generous help

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