Escherichia coli’de Metallere ve pH Stresine karşı RpoS’nin Rolünün Araştırılması

Year 2018, Volume: 7 Issue: 2, 105 - 121, 17.08.2018

Cihan Darcan , Özge Kaygusuz Ezgi Aydın

https://doi.org/10.18036/aubtdc.372141

Abstract

Escherichia coli’de
alternatif sigma faktörü olarak adlandırılan rpoS (σ³⁸) asit şoku, açlık stresi, ozmotik stres,
oksidatif stres, DNA hasarı ve durağan faza geçiş dahil olmak üzere farklı streslere
karşı hücresel cevapta görevli olan genlerin ekspresyonunu kontrol etmektedir. Bu
çalışmada Escherichia coli W3110’da rpoS geninin metal ve pH stresi altındaki
rolü araştırılmıştır. E. coli BW25113
suşunda mutant olan rpoS geni, P1kc
fajı ile yabani tip E. coli W3110’a
aktarılmıştır. Yabani tip E. coli
W3110 ve rpoS mutant E. coli W3110 suşunda yaşam deneyleri pH
5.5, 7.0 ve 8.0 fosfat tamponunda olmak üzere 3 farklı pH’da plak sayım metodu
ile yapılmıştır. 6 farklı metal varlığında (Zn, Ni, Co, Cd, Ag ve Cu) metal
stresindeki rolü minimal inhibisyon konsantrasyonu ve petri damlatma yöntemi
ile belirlenmiştir. Çalışılan tüm pH değerlerinde rpoS mutant suşlarda duyarlılık gözlenmiştir. Cd metaline karşı rpoS mutantının oldukça önemli bir
direnç gösterdiği belirlenirken, diğer metallerde herhangi bir rolü olmadığı
tespit edilmiştir. rpoS geninin rolü,
genin tamamlanması yapılarak doğrulaması sağlanmıştır. Bu sonuçlar RpoS
alternatif sigma faktörünün Cd metali ile ilişkili genlerin kontrolünde ve pH
stresi ile ilişkili genlerde rolü olduğunu göstermektedir

Keywords

Escherichia coli, rpoS, Metal Stresi, Kadmiyum

METALLERE VE pH STRESİNE KARŞI Escherichia coli’de RpoS’NİN ROLÜNÜN ARAŞTIRILMASI

Abstract

RpoS (σ38), which
acts as an alternative sigma factor in Escherichia
coli, controls the expression of genes responsible for cellular responses
to different stresses including acidity, starvation, osmotic, oxidative stress,
DNA damage, and stationary phase transition. In this study, the role of the rpoS gene in Escherichia coli
W3110 under metals and pH stress was researched. The rpoS gene, which is a mutant in the E. coli BW25113 strain, was transferred by transduction with the
P1kc phage to wild-type E. coli
W3110. Life experiments in wild-type E.
coli W3110 and rpoS mutant E. coli W3110 strains were carried out
by plaque counting method at 3 different pH, in pH 5.5, 7.0 and 8.0 phosphate
buffer. The role of rpoS in the
presence of 6 different metals (Zn, Ni, Co, Cd, Ag and Cu) was also determined
by the method of dropping petri. Sensitivity at rpoS mutant strains was
observed at all pH values studied. It has been determined that the rpoS mutant
E. coli against Cd metal shows a very significant resistance, but it has been
found that there is no role in other metals. Verification of the role of the
rpoS gene has been achieved by genetic complementation. These results show that
the RpoS alternative sigma factor plays a role in the control of genes related
to the Cd metal and in the genes associated with pH stress.

Keywords

Escherichia coli, rpoS, Metal Stress, cadmium

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