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The activity of eugenol antibiotic on E. coli: Studies to identify the genes involved in the mechanism of action of the antibiotic
Başlık:
The activity of eugenol antibiotic on E. coli: Studies to identify the genes involved in the mechanism of action of the antibiotic
Yazar:
Alkhaibari, Noura Saler, author.
ISBN:
9780438078932
Yazar Ek Girişi:
Fiziksel Tanımlama:
1 electronic resource (39 pages)
Genel Not:
Source: Masters Abstracts International, Volume: 57-06M(E).
Advisors: Mrinal Bhattacharjee.
Özet:
The genus Escherichia coli is named after Theodor Escherich, who isolated this type species of the genus in 1885. Escherichia coli is a gram-negative bacteria, facultatively anaerobic, rod-shaped, fermenter, and is oxidase negative. Large numbers of this bacteria are normally found in the gastrointestinal tract, thus those bacteria might be opportunistic when the intestine's surface is perforated and the organism penetrates the peritoneal cavity. In addition; Escherichia coli that are part of normal flora have the ability to create infection when the defenses of patient are compromised.
Eugenol, also known as clove oil, is an essential oil that is extracted from the clove bud. It commonly used as anesthesia in oral infection, analgesic, spice, flavor, and in other industrial applications. It also has anti-inflammatory properties. It can affect both Gram-negative and Gram-positive bacteria. Microorganisms can be resistant to some antibiotic agents due to the effective permeability barrier function of membrane. However, eugenol oil can denature the proteins and react with the cell membrane phospholipids thereby changing the permeability.
Some essential oils have activity against bacteria. The activity of eugenol oil antibiotic on E. coli was tested by zone of inhibition. We have identified the minimum inhibitory concentration (MIC) on LB plate and broth. Then, eugenol resistant mutants were obtained by passages in LB broth containing increasing concentrations of eugenol, and colonies were saved as eugenol resistant mutants. The plasmid pVJT128 was transferred to the one of mutant strain, LIU185 by conjugation method to obtain the strain L1U198 [MV10Nal, Eug (pVJT128)]. The transposase gene present in pVJT128 was induced by using IPTG to obtain a library of random transposon insertion mutants which were selected on kanamycin plates. The library of mutants was replica plated from Km plate to eugenol plate to identify revertant colonies that were resistant to eugenol but sensitive to kanamycin. The genomic DNA for those revertants was isolated by phenol extraction to identify the gene that contain the transposon insertion. Then, the DNA was digested by BamHI and BglII, ligated, and amplified by inverse PCR. In order to get the band for the insertion, the DNA for the revertant and plasmid pMB78 were digested using BamHI and then ligated the DNA with pMB78, which were then transformed to E. coli strain by CaCl2 method. After that the plasmid DNA was isolated form a transformant for further studies to identify the gene that is involved in the mechanism of action of eugenol.
Notlar:
School code: 0198
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Yer Numarası | Demirbaş Numarası | Shelf Location | Lokasyon / Statüsü / İade Tarihi |
---|---|---|---|
XX(696236.1) | 696236-1001 | Proquest E-Tez Koleksiyonu | Arıyor... |
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