Bazı Kimyasalların Etkisi Altında Escherichia coli’nin OmpF - OmpC Porin Protein Sentezi ve Bu Sentezde EnvZ, RpoS, H-NS, AcP’ın Rolünün Belirlenmesi

Öz

Escherichia coli’nin dış membranında bulunan OmpC ve OmpF porin proteinleri, stres koşullarına karşı korunmasında önemli yer tutar. Bu çalışmada dezenfektan özellikli çeşitli kimyasal maddelerin E. coli’nin OmpC ve OmpF porin protein sentezini nasıl etkilediği araştırılmıştır. E. coli’de, formaldehit, klor, etanol, çamaşır suyu, H₂O₂ ve Sodyum Dodesil Sülfatın (SDS) belirlenen konsatrasyonları besiyerine eklenerek, ompC ve ompF genleri yerine raportör gen olarak eklenen β-galaktosidaz’ın aktivitesi spektrofotometrik olarak ölçülmüş ve elde edilen sonuçlar kontrol grupları ile karşılaştırılmıştır. Sonuç olarak çalışılan kimyasal maddelerin E. coli’nin büyümesini oldukça yavaşlattığı, ancak yinede üremenin gerçekleştiği belirlenmiştir. E. coli’de OmpF sentezinin formaldehit, etanol ve SDS ilave edildikten 2 saat sonra oldukça azaldığı, çamaşır suyu ve klor ilavesinde biraz azaldığı ve H₂O₂ ilave edildiği zaman ise değişmediği tespit edilmiştir. OmpC sentezinde ise klor, formaldehit, H₂O₂ ilave edildiğinde %50 oranında azaldığı belirlenirken, çamaşır suyunda yaklaşık 2 kat ve SDS ilave edilen örneklerde ise 1.5 katlık bir artış belirlenmiştir. Ayrıca hem OmpC hem de OmpF porin proteininin sentezinde RpoS, Pta, EnvZ ve H-NS proteinlerinin rollerinin olduğu belirlenmiştir. Ancak moleküler mekanizmaları henüz aydınlatılamamıştır.

Anahtar Kelimeler

• Porin,Dezenfektan kimyasallar,Escherichia coli,Porin

The Role of EnvZ, RpoS, H-NS, AcP and Synthesis of OmpC-OmpF Porin Proteins of Escherichia coli Under Effect of Some Chemicals

Öz

OmpC and OmpF porin proteins, located on the outer membrane of Escherichia coli, play an important role in protection against stress conditions. In this study, it was investigated how various chemical substances as a disinfectant affect E. coli OmpC and OmpF porin protein synthesis. The activity of β-galactosidase, added as a reporter gene in place of ompC and ompF genes, was measured spectrophotometrically by adding formaldehyde, chlorine, ethanol, bleach, H₂O₂ and Sodium Dodecyl Sulfate (SDS), used as disinfectants in E. coli. The results were compared with control groups. As a result, it was determined that the studied chemical substances slowed down the growth of E. coli at the added concentrations, but continued to multiply. It was determined that the OmpF synthesis in E. coli decreased considerably after 2 hours from the addition of formaldehyde, ethanol and SDS, decreased slightly during the addition of bleach and chlorine, and did not change when H₂O₂ was added. In the OmpC synthesis, when chlorine, formaldehyde, and H₂O₂ were added, it was determined to decrease by 50% compared to the untreated state, while the increase was found as a 1.5 time in the cases where SDS was added and about 2 times in the bleach. In these alterations, it has been determined that the roles of RpoS, Pta, EnvZ, and H-NS proteins on both OmpC and OmpF porin protein synthesis but have not yet been elucidated by their molecular mechanisms.

Anahtar Kelimeler

• Disinfectan chemicals,Escherichia coli,Porin

Kaynakça

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