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Konjugacija plazmida pOX38:Cm v izbrane seve bakterije Escherichia coli
ID Repar, Polona-Maja (Author), ID Starčič Erjavec, Marjanca (Mentor) More about this mentor... This link opens in a new window, ID Kastrin, Andrej (Co-mentor)

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Abstract
Povečevanje števila proti antibiotikom odpornih bakterij postaja vedno večji problem. Geni za odpornost so pogosto zapisani na mobilnih genetskih elementih, ki se lahko s horizontalnim genskim prenosom hitro prenašajo med bakterijami. Med horizontalnimi genskimi prenosi velja konjugacija, genski prenos ob neposrednem stiku dveh bakterij, kot najpomembnejši dejavnik za širjenje odpornih bakterij. Dobro poznavanje dejavnikov, ki vplivajo na frekvenco konjugacije, lahko vodi v odkritje novih strategij za borbo proti odpornim bakterijam. Med te dejavnike spadajo obrambni mehanizmi bakterij pred vdorom tuje DNA, kot sta na primer sistem CRISPR-Cas in restrikcijsko-modifikacijski (R-M) sistem. Cilj magistrskega dela je bil preveriti, ali obstaja povezava med frekvenco konjugacije plazmida pOX38:Cm in prisotnostjo genskih zapisov za sistem CRISPR-Cas in R-M sisteme v recipientskih sevih, inkompatibilnostjo plazmidov recipientskih in donorskih sevov ali pa s protimikrobnim delovanjem recipientskih sevov. CRISPR-Cas in R-M sisteme smo v posameznih sevih poiskali s prosto dostopnima računalniškima programoma (CRISPR-Cas++ in REBASE), vendar nismo dokazali nobene povezave s frekvenco konjugacije. Nato smo preverili še sposobnost recipienta za zaviranje rasti donorja (test za prisotnost bakteriocinov) in s programom BLAST preverili inkompatibilnost plazmidov v recipientskih sevih s plazmidom pOX38:Cm. Nekateri recipientski sevi so sicer povzročili cono lize donorskega seva HB101 pOX38:Cm, a povezava s frekvenco konjugacije ni obstajala. Primerjava nukleotidnih zaporedij pOX38:Cm in plazmidov recipientskih sevov tudi ni razkrila morebitne inkompatibilnosti med plazmidi. Opaženih razlik v frekvencah konjugacije pOX38:Cm v različne recipientske seve tako nismo mogli povezati ne z sistemom CRISPR-Cas, ne R-M sistemi, ne protimikrobnim delovanjem in tudi ne z inkompatibilnostjo plazmidov.

Language:Slovenian
Keywords:plazmidi, frekvenca konjugacije, pOX38:Cm, CRISPR-Cas, restrikcijsko-modifikacijski sistem, inkompatibilnost plazmidov, protimikrobno delovanje
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[P.-M. Repar]
Year:2021
PID:20.500.12556/RUL-130413 This link opens in a new window
UDC:579.25:577.2.083
COBISS.SI-ID:76675843 This link opens in a new window
Publication date in RUL:15.09.2021
Views:914
Downloads:99
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Secondary language

Language:English
Title:Conjugation of the pOX38:Cm plasmid into selected bacterial strains of Escherichia coli
Abstract:
The growing number of antibiotic resistant bacteria is becoming an increasing problem. The genes for resistance are often carried on mobile genetic elements that can spread rapidly through bacterial populations by horizontal gene transfer. Mechanisms of horizontal gene transfer include conjugation, the gene transfer between two bacteria that are in physical contact, which is considered the most important factor in the spread of resistant bacteria. A good characterisation of the factors that influence the conjugation frequency may lead to the discovery of new strategies to combat the resistant bacteria. These factors include bacterial defence mechanisms to control the entry of foreign DNA, such as CRISPR-Cas and restriction-modification (R-M) systems. The aim of this Master Theses was to determine whether there is a correlation between conjugation frequency of the plasmid pOX38:Cm and the presence of genes for CRISPR-Cas and R-M systems in recipient strains, incompatibility of plasmids in recipient and donor cell or antimicrobial production of recipient strains. CRISPR-Cas and R-M systems were determined using freely available computer programs (CRISPR-Cas++ and REBASE), but a correlation with the conjugation frequency was not discovered. Therefore we investigated the ability of the recipient to inhibit the growth of the donor (bacteriocinogenity assay) and the incompatibility between the plasmids in recipient cells and plasmid pOX38:Cm (BLAST). Some recipient strains formed lysis zones of the donor strain, but a correlation with the conjugation frequency did not exist. Comparison of the plasmids revealed no incompatibility between them. The observed differences in conjugation frequencies could not be explained by CRISPR-Cas or R-M systems, antimicrobial production or plasmid incompatibility.

Keywords:plasmids, conjugation frequency, pOX38:Cm, CRISPR-Cas, restriction-modification system, plasmid incompatibility, antimicrobial production

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