TY - JOUR JF - Iran-J-Med-Microbiol JO - Iran J Med Microbiol VL - 7 IS - 4 PY - 2014 Y1 - 2014/3/01 TI - Recombinant Streptomyces clavuligerus strain including cas2 gene production and analysis its antibiotic overproduction by bioassay TT - تولید گونه‌ی تراریخت استرپتومایسس کلاولی‌جروس واجد ژن cas2 و بررسی افزایش تولید پادزیست در آن با روش بیواسی N2 - Background: Streptomyces clavuligerus is one of the most important strain that produce clavulanic acid that wildly used in combination of strong but sensitive to β-lactamase antibiotics in clinics. The cas2 is one of the important genes in the biosynthesis pathway of clavulanic acid. Materials and Methods: The recombinant construct pMTcas2 which contain cas2 gene is obtained from Isfahan University. Recombinant plasmid extracts from streptomyces lividans and confirm by enzyme digestion. The streptomyces clavuligerus protoplast was prepared and transformation was done by using polyethylene glycol. Transformation was confirmed by plasmid extraction and PCR using cas2 specific primers. Finally, bioassay method was used to survey the effect of extra copy of cas2 on clavulanic acid production. Result: Plasmid extraction was initially carried out and the structure of plasmid was confirmed by digestion. The typical white colony was seen on protoplast recovery culture containing thiostrepton antibiotic and gray spores were detected after one week. Plasmid extraction was done from transformed strain and transformation was confirmed by PCR. The results of the bioassay show that amplification of the cas2 gene in multicopy plasmids resulted in a 4.1 fold increase in clavulanic acid production. Conclusion: The bioassay was done and the diameters of zone of inhibition in control and sample were compared. The results of the bioassay show that amplification of the cas2 gene in multicopy plasmids resulted in a 4.1 fold increase in clavulanic acid production. Overproduction of clavulanic acid decreases the cost of its dependent drug production. SP - 30 EP - 35 AU - Hojati, Zohreh AU - Givi, Sedighe AU - Kay, Maryam AD - University of Isfahan KW - Streptomyces clavuligerus KW - clavulanic acid KW - Bioassay KW - pMTcas2 plasmid KW - cas2 gene. UR - http://ijmm.ir/article-1-193-en.html ER -