master's thesis
ISPITIVANJE SVOJSTAVA BAKTERIJE LEGIONELLA LONGBEACHAE OBILJEŽENE ZELENO FLUORESCIRAJUĆIM PROTEINOM

Anamaria Trbović (2015)
Sveučilište u Rijeci
Medicinski fakultet
Katedra za mikrobiologiju i parazitologiju
Metadata
TitleISPITIVANJE SVOJSTAVA BAKTERIJE LEGIONELLA LONGBEACHAE OBILJEŽENE ZELENO FLUORESCIRAJUĆIM PROTEINOM
AuthorAnamaria Trbović
Mentor(s)Ivana Gobin (thesis advisor)
Abstract
Legionella longbeachae je gram-negativni štapić koji je kompliciran za genetsku manipulaciju te je stoga malo poznato o ţivotnom ciklusu ove bakterije. Kako bismo doprinjeli ispitivanju patogeneze žţivotnog ciklusa L. longbeachae, cilj ovog rada bio je proizvesti bakteriju obiljeţenu zeleno fluorescirajućim proteinom te ispitati njezina svojstva. U istraţivanju su korištena dva soja bakterije L. longbeachae: soj NSW 150 i soj A5H5. U pokusima je korišten plazmid pMip koje je sadržavao gen gfp te gen za rezistenciju na kloramfenikol, a korištena metoda unosa plazmida u bakterijsku stanicu bila je elektroporacija. Ispoljavanje gfp gena praćeno je pod fluorescentnim mikroskopom. Nadalje se ispitalo postoji li razlika u brzini rasta u AYE bujonu izmeĎu legionela koje su obiljeţene GFP-om i neobiljeţenih legionela. S obzirom da se amebe često koriste kao modeli za istraţivanje unutarstaničnog ţivotnog ciklusa legionele ispitivala se funkcionalnost obiljeţenih bakterija kokultivacijom s amebom Acanthamoeba castellanii te se pratio broj bakterija tijekom kokultivacije. Rezultati su pokazali da ispitivani sojevi fluoresciraju zeleno u odnosu na kontrolu te da nema razlike u brzini rasta oba soja bakterije. Rezultati takoĎer pokazuju da je došlo do porasta broja bakterija tijekom kokultivacije sa amebama te da nema značajne razlike izmeĎu ispitivanih sojeva legionela. Za bakteriju L. longbeachae obilježenu GFP-om smo u ovom radu dokazali, da se s obzirom na ispitivana svojstva, ponaša kao divlji soj te se stoga ova mutanta moţe koristi i u daljnim pokusima za ispitivanje ţivotnog ciklusa i patogenosti L. longbeachae.
KeywordsLegionella green fluorescent protein (GFP) amoeba mutant
GranterSveučilište u Rijeci
Medicinski fakultet
Lower level organizational unitsKatedra za mikrobiologiju i parazitologiju
PlaceRijeka
StateCroatia
Scientific field, discipline, subdisciplineBIOMEDICINE AND HEALTHCARE
Clinical Medical Sciences
Medical Microbiology
Study programme typeuniversity
Study levelgraduate
Study programmeStudy of Sanitary Engineering
Academic title abbreviationmag. sanit. ing.
Genremaster's thesis
Language Croatian
Defense date2015
Parallel abstract (English)
Legionella longbeachae is a gram-negative bacili difficult for genetic manipulation and because of which is very little known about it's life cycle. In order to contribute to study od life cycle and pathogenesis of L. longbeachae, the aim of this study was to produce a bacteria expressing green fluorescent protein and test its properties. In this study 2 strains of bacteria L. longbeachae: NSW 150 and A5H5 was used. In the experiments a plasmid pMip that contained a gen gfp as well a gen for resistance on chloramphenicol was used and the method was electroporation. Expression of gfp gene was monitored by fluorescent microscop. In addition, we tested whether there is a difference between the marked and unmarked legionela in the growth in AYEmedium . Since amoebas are often used as models for studying the intracellular life of legionela, the functionality of marked bacteria in co-cultivation with amoeba Acanthamoeba castellanii was tested and the number of bacteria was monitored during the cultivation. The results showed that the marked strains have the ability to fluorescent green in contrast with wild type strains, moreover therewas no difference in growth rate of either strain of bacteria. Results also showed that there has been an increase in the number of bacteria during the cultivation with amoeba, and that there is no significant difference between tested strains. In this study we showed that L. longbeachae expressing GFP, in regards of tested proterties, acts as wild type strains and therefore it can be used in further experiments to test life cycle and pathogenecity of L. longbeachae.
Parallel keywords (Croatian)Legionella zeleno fluorescirajući protein (GFP) ameba mutanta
Resource typetext
Access conditionAccess restricted to students and staff of home institution
Terms of usehttp://rightsstatements.org/vocab/InC/1.0/
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:184:239026
CommitterBosa Licul