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Exploring the role of Bacillus subtilis gene kamA controlled by a phage regulatory switch
ID Kračman, Filip (Author), ID Dragoš, Anna Magdalena (Mentor) More about this mentor... This link opens in a new window

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Abstract
Bacillus subtilis is a model organism for studying Gram-positive microorganisms; its kamA gene can be interrupted by the inserted lysogenic phage phi3T, closely related to SPbeta which was shown to act as a phage regulatory switch regulating the expression of the spsM gene during sporulation. This thesis explores the possible antiviral activity of the kamA gene product based on similarity of its 3D structure with three viperins, a class of antiviral proteins found across the tree of life. This possible role was tested by performing multiple double-layer plaque assays, susceptibility assays in liquid cultures and a phage adsorption assay comparing B. subtilis strain PY79 and its ∆kamA and ∆spsM mutants. Results revealed that a deletion of kamA decreases susceptibility of B. subtilis to lytic phage Nf, rather than the hypothesised viperin-like activity of its gene product. Furthermore, deletion of the gene exhibits an effect on the rate of phage adsorption resulting in a trend distinct from cells with an intact gene, However, due to an unforeseen phenomenon affecting this experiment, we cannot say this with certainty. As kamA harbours an attachment site for phage phi3T which effectively has a deleterious effect, these findings demonstrate a way in which lysogenic phages may confer protection towards lytic phages.

Language:English
Keywords:lysogenic bacteriophages, phage regulatory-switch, bacteriophage resistance, Bacillus subtilis, gene kamA
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[F. Kračman]
Year:2024
PID:20.500.12556/RUL-163849 This link opens in a new window
UDC:579.25:578.347
COBISS.SI-ID:211345155 This link opens in a new window
Publication date in RUL:13.10.2024
Views:84
Downloads:33
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Secondary language

Language:Slovenian
Title:Proučevanje uravnave gena kamA bakterije Bacillus subtilis s fagnim regulatornim stikalom
Abstract:
Gen kamA bakterije Bacillus subtilis, ki je modelni organizem za preučevanje grampozitivnih mikroorganizmov, je lahko prekinjen z vstavljenim lizogenim fagom phi3T, ki je soroden fagu Spbeta. Zanj je bilo dokazano, da deluje kot regulatorno fagno stikalo, ki uravnava izražanje gena spsM med sporulacijo. V tej magistrski nalogi smo preučili možno protivirusno aktivnost produkta gena kamA. Slednja aktivnost proteina je bila predvidena na podlagi podobnosti 3D modela strukture proteina KamA s strukturami treh protivirusnih proteinov, viperinov, ki jih lahko najdemo po vsem drevesu življenja. Morebitno protivirusno vlogo gena kamA smo preizkusili z več občutljivostnimi testi tako na ploščah kot v kulturi, ter s spremljanjem hitrosti adsorpcije litičnega faga Nf na gostitelja, in sicer tako, da smo primerjali vpliv faga na sev B. subtilis PY79 ter njegovi ∆kamA in ∆spsM mutanti. Rezultati so pokazali, da delecija gena kamA zmanjša občutljivost B. subtilis na fag Nf, vendar pa odpornosti ne daje domnevana viperinu-podobna aktivnost proteina KamA. Rezultati nakazujejo, da delecija gena kamA vpliva na hitrost adsorpcije faga Nf, ki se razlikuje od adsorpcije na celice divjega tipa. Vendar zaradi nepredvidenega pojava, ki je vplival na ta poskus tega ne moremo trditi dokončno. Ker lahko fag phi3T gen prekine z vstavitvijo svojega genoma v ta gen, te ugotovitve nakazujejo način, s katerim lizogeni fagi lahko vplivajo na svojega gostitelja in kako mu zagotovijo zaščito pred drugimi fagi.

Keywords:lizogeni bakteriofagi, fagno regulatorno stikalo, odpornost na bakteriofage, Bacillus subtilis, gen kamA

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