İnsulin ve glukozun E. coli’de üreme ve mar regulonunun ekspresyonu üzerine etkisi

Year 2019, , 209 - 216, 12.06.2019

Defne Gümüş , Fatma Kalaycı Yüksek , Gülşen Uz Aslı Bayraktar Pınar Köseoğlu Yağmur Ofluoğlu Benay Daylan Mine Anğ Küçüker

https://doi.org/10.18663/tjcl.459417

Abstract

Amaç:Konak
faktörlerinin bakterilerin virulans, antibiyotik duyarlılığı ve üreme gibi
özelliklerini etkilediği bilinmektedir. Bu çalışmada insan insulin ve glukozunE. coli SPC105 suşunun üremesi ve
marregulonunun (marA, marR, ompF, acrA, acrB vetolC genlerini içeren) ekspresyonu üzerine etkisinin araştırılması
hedeflenmiştir.

Gereç ve Yöntemler:E. coli SPC105 triptik soy buyyonda
(TSB-kontrol) ve 20 µU/mLinsulin, 200 µU/mLinsulin, %0,1 glukoz ve 200
µU/mLinsulin + %0,1 glukoz içeren TSB’de üretilmiştir. Üremeler türbidimetrik
yöntem ile belirlenmiştir. Gen ekspresyonu düzeyleri kantitatifpolimeraz zincir
reaksiyonu (qPCR) ile araştırılmıştır. İstatistiksel analizlerde Tukey’in post
hoc-testi kullanılmıştır.

Bulgular: Absorbans
sonuçlarına göre %0,1 glukoz ve 200 µU/mLinsulin + %0,1 glukoz içeren
besiyerleri üremeyi anlamlı düzeyde düşürmüştür. acrA geninin ekspresyon seviyesinin 200µU/mL insülin içeren
besiyerinde düştüğü görülmüştür. marR
ve marA’nın ekspresyonları da benzer
şekilde 200 µU/mLinsulin, %0,1 glukoz ve 200 µU/mLinsulin + %0,1 glukoz içeren
TSB’de azalmıştır. marA, marR, acrA ekspresyonlarındaki azalış istatistiksel olarak anlamlı
bulunmuştur (p<0.05).

Sonuç: Bu çalışmanın sonuçları bir kez daha
göstermiştir ki, konak faktörleri bir bakterinin üremesi üzerine olduğu kadar
antibiyotik direnci ile ilişkili farklı genlerinde ekspresyonu üzerinde etkili
olabilmektedir. 

Keywords

glukoz, insulin, mar operon, gen ekspresyonu, üreme

The effects of insulin and glucose on growth and expression of mar regulon in E. coli

Abstract

Aim:Host factors
are known to modulate virulence, antibiotic susceptibility and growth of
bacteria. In the present study we aimed to investigate the effect of human
insulin and glucose on growth and expression of mar regulon (marA, marR, ompF, acrA, acrB and tolC) of E. coli SPC105.

Material and Methods:E. coli SPC105 was
grown in tryptic soy broth (TSB-control) and TSB containing 20 µU/mL insulin,
200 µU/mL insulin, 0.1% glucose and 200 µU/mL insulin + 0.1% glucose. Growths
were determined via turbidimetric method. 
Quantitative polymerase chain reaction (qPCR) was used to determine the
gene expression levels. Statistical analysis were performed via Tukey’s post
hoc-test.

Results:According to
absorbance values, it has been shown that 0.1% glucose and 200 µU/mL insulin +
0.1% glucose have led a significant decrease on growth. Expression of acrA gene was shown to be decreased in
the presence of 200μIU insulin. Expressions of marR and marA were also
decreased in 200μU insulin, 0.1 % glucose+200μU insulin and 0.1% glucose added
TSB. Decreasing of marA, marR and acrA expressions were shown to be statistically significant
(p<0.05).

Conclusion:The results
of the present study has shown once more that host factors may influence the
growth of a bacterium as well as gene expressions associated with antibiotic
susceptibilities.

Keywords

glucose, insulin, mar operon, gene expression, growth

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