Priprava proti ciprofloksacinu odpornega seva N4i bakterije Escherichia coli

magistrsko delo

Petra Drčar (Author), Marjanca Starčič Erjavec (Mentor), Andrej Kastrin (Mentor)

Abstract

Zaradi naraščanja števila proti antibiotikom odpornih bakterij, se vedno bolj izraža potreba po alternativnih protimikrobnih učinkovinah. Ena izmed njih je konjugativni plazmid pOX38a z zapisom za kolicin E7. Za preverjanje njegove učinkovitosti smo v okviru magistrske naloge s transdukcijo želeli pripraviti proti ciprofloxacinu odporen sev N4i bakterije Escherichia coli. S karakterizacijo (plazmidna izolacija, konjugacija na plošči in preverjanje bakteriocinske aktivnosti ) naravnih uropatogenih, proti ciprofloksacinu odpornih sevov DL bakterije E. coli, smo izbrali sev, iz katerega smo v več poskusih transducirali zapis odpornosti proti ciprofloxacinu v sev N4i. Ker s transdukcijo nismo uspeli prenesti zapisa, smo proti ciprofloksacinu odporen sev N4i, selekcionirali s precepljanjem v gojiščih z naraščajočo koncentracijo ciprofloksacina. Genom pridobljenega seva N4i smo sekvencirali z napravo GridIon. Pridobljeni podatki so bili obdelani s pripadajočo programsko opremo MinKNOW, sledilo je demultipleksiranje in identifikacija nukleotidnih baz iz surovih podatkov, z uporabo programa Guppy. Sestavljanje genoma de novo je potekalo s pomočjo algoritma Flye. Sestavljeno nukleotidno zaporedje smo primerjali z zaporedjem seva Nissle 1917, ki je dostopno v podatkovni bazi NCBI. Preverili smo tudi ali so ugotovljene razlike v nukleotidnem zaporedju locirane v že znanih genih, povezanih z odpornostjo proti fluorokinolonskim antibiotikom.

Keywords

antibiotiki;odpornost;ciprofloksacin;alternativa;Escherichia coli;magistrska dela;

Data

Language:	Slovenian
Year of publishing:	2022
Typology:	2.09 - Master's Thesis
Organization:	UL BF - Biotechnical Faculty
Publisher:	[P. Drčar]
UDC:	579
COBISS:	122149891
Views:	15
Downloads:	2
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Preparation of a ciprofloxacin-resistant N4i Escherichia coli strain
Secondary abstract:	As the number of bacteria resistant to antimicrobial drugs is increasing, the need for alternative antimicrobial agents is becoming more apparent. The conjugative plasmid pOX38a carrying the colicin E7 gene is an alternative antimicrobial. In order to test its efficiency, a ciprofloxacin resistant Escherichia coli N4i strain was needed, hence the goal of this thesis was to prepare such a strain. After initial characterization (plasmid isolation, mating assay on solid media, and bacteriocin production) of natural uropathogenic ciprofloxacin-resistant E. coli DL strains, which were a possible source of the ciprofloxacin resistance gene, we started to perform different transduction experiments in order to transfer the ciprofloxacin resistance gene into N4i. Since desipite several attemps we failed to transfer ciprofloxacin resistance gene with transduction, the ciprofloxacin-resistant bacterial strain N4i was grown by stepwise cultivation in media with increasing concentrations of ciprofloxacin. The genome of the obtained strain was sequenced using a GridIon instrument. The data obtained were processed using the appropriate MinKNOW software, followed by demultiplexing and identification of bases from the raw data using Guppy program. The genome was assembled de novo using the Flye algorithm. The assembled nucleotide sequence was compared with the deposited nucleotide sequence of strain Nissle 1917 in the NCBI database and analyzed, if the observed nucleotide differences were located in already known genes associated with resistance to fluoroquinolone antibiotics.
Secondary keywords:	antibiotics;resistence;ciprofloxacin;alternative;master thesis;Mikrobiologija;Univerzitetna in visokošolska dela;
Type (COBISS):	Master's thesis/paper
Study programme:	0
Embargo end date (OpenAIRE):	1970-01-01
Thesis comment:	Univ. v Ljubljani, Biotehniška fak., Oddelek za biologijo
Pages:	1 spletni vir (1 datoteka PDF (XII, 68 str., [1] str. pril.)
ID:	16479286