Integrative gene cloning and expression system for Streptomyces sp. US 24 and Streptomyces sp. TN 58 bioactive molecule producing strains.
Streptomyces sp. US 24 and Streptomyces sp. TN 58, two strains producing interesting bioactive molecules, were successfully transformed using E. coli ET12567 (pUZ8002), as a conjugal donor, carrying the integrative plasmid pSET152. For the Streptomyces sp. US 24 strain, two copies of this plasmid were tandemly integrated in the chromosome, whereas for Streptomyces sp. TN 58, the integration was in single copy at the attB site. Plasmid pSET152 was inherited every time for all analysed Streptomyces sp. US 24 and Streptomyces sp. TN 58 exconjugants under nonselective conditions. The growth, morphological differentiation, and active molecules production of all studied pSET152 integrated exconjugants were identical to those of wild type strains. Consequently, conjugal transfer using pSET152 integration system is a suitable means of genes transfer and expression for both studied strains. To validate the above gene transfer system, the glucose isomerase gene (xylA) from Streptomyces sp. SK was expressed in strain Streptomyces sp. TN 58. Obtained results indicated that heterologous glucose isomerase could be expressed and folded effectively. Glucose isomerase activity of the constructed TN 58 recombinant strain is of about eighteenfold higher than that of the Streptomyces sp. SK strain. Such results are certainly of importance due to the potential use of improved strains in biotechnological process for the production of high-fructose syrup from starch.
Sioud
Samiha
S
Laboratory of Prokaryotic Enzymes and Metabolites, Centre of Biotechnology of Sfax, Road of Sidi Mansour Km 6, P. O. Box 1177, 3018 Sfax, Tunisia.
Aigle
Bertrand
B
Karray-Rebai
Ines
I
Smaoui
Slim
S
Bejar
Samir
S
Mellouli
Lotfi
L
D019747:Aldose-Ketose Isomerases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001287:Attachment Sites, Microbiological / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D001483:Base Sequence; D050260:Carbohydrate Metabolism / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D018014:Gene Transfer Techniques; D016982:Micrococcus luteus / Q000187:drug effects; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D016415:Sequence Alignment; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase