Doğal ortamlardan izole edilen alkalifilik toprak bakterilerinde plasmid kodlu na-tellurit dirençliliği ve transformasyon olanaklarının araştırılması

Bu çalışmada Na-tellurit bileşiğinin toprak bakterilerinden alkalifilik Bacillus spp. suşlarında inhibitör etkisi araştırıldı.Bakteriler, Na-tellurit bileşiğine karşı 1-180 μg/ml artan konsantrasyonlarda denendi. Na-tellurit bileşiğinin 1-60 μg/mlkonsantrasyonlardaki denemesinde izolatların çoğunluğunun ürediği gözlendi. 70-180 μg/ml olarak denenen konsantras yonlarda ise hassasiyet profilinin arttığı saptandı. 180 μg/ml konsantrasyonlarda ise % 100 hassasiyet gözlendi. Ayrıca,izolatlar için MICs değeri 0.8 μg/ml olarak belirlendi. İzolatların Na-tellurit bileşiğine karşı plasmid kodlu dirençliliğininsaptanması için acridine orange (20 μg/ml) ve ethidium bromid (10-5M) ile plasmid eliminasyonu yapıldı. İzolatlar’daBacillus spp.’ye ait % 60.86 Na-tellurit bileşiğine hassasiyet, % 39.13 dirençlilik fenomeni ve % 21.10 oranında da plasmidkodlu Na-tellurit dirençliliği bulundu. Na-tellurit bileşiğine ve antibiyotiğe karşı dirençlilik genini taşıyan plasmidler (B. cereusT112-Na-TeR), Na-tellurit bileşiğine ve antibiyotiğe hassasiyet gösteren suşa (B. megaterium DSM 32-Na-TeS) protoplasttransformasyonu yoluyla aktarıldı ve 108 transformant elde edildi. Agaroz jel elektroforez seperasyonu sonucunda plasmidin22.514 bp moleküler ağırlığa sahip olduğu saptandı.

Investigations upon the likelihood of transformation and resistance of na-tellurite originated plasmide at the soil bacteria ısolated from natural environment

In this study, the inhibitory effects of Na-tellurit compound at the Alkalophilic Bacillus spp. strains isolated fromthe soil bacteria were investigated. Bacteria were tested against the increasing concentrations of Na-tellurite at 1-180μg/ml. The isolates of Na-tellurite at the 1-60 μg/ml concentrations mostly showed resistance, but sensitivity wasdetermined at 70-180 μg/ml. Complete (100 %) sensitivity was observed at 180 μg/ml. For Bacillus spp., the MICs of Na tellurit were found to be 0.8 μg. Plasmid elimination was made using acridine orange (20 μg/ml) and ethidium bromide (10-5 M) for determination of the isolate resistance originated from plasmid against Na-tellurite. In the isolates of Bacillus spp,there were 60.86 % sensitivity, 39.13 % resistance phenomenon and 21.10 % ratio of resistance originated from plasmidagainst Na-tellurite. Plasmids of B. cereus T112-Na-Te R, carrying the resistance gene against the antibiotic and Na-tellurite,were transferred by protoplast transformation to B. megaterium DSM 32-Na-Te S strain showing sensitivity against antibioticand Na-tellurit. A number of 108transformants from the soil isolates were obtained by protoplast transformation. Theseperation of plasmid was made by agarous gel electrophoresis and, finally it was determined that the plasmid had 22,514bp of molecular weight.

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