diplomsko delo
Kim Glavič (Author), Marko Dolinar (Mentor)

Abstract

Sistemi toksin-antitoksin (TA) so genetski elementi, ki zapisujejo za relativno stabilen proteinski toksin, ki onemogoči rast celice, in nestabilen antitoksin, ki zavre škodljivi učinek kompatibilnega toksina. Njihova biološka vloga še ni povsem razumljena, znano pa je, da sodelujejo pri mnogih bioloških procesih, izmed katerih izstopajo postsegregacijska smrt, abortivna infekcija in bakterijska persistenca. Najdemo jih na številnih plazmidih in kromosomih mnogih arhej ter bakterij, med katere sodi tudi cianobakterija Microcystis aeruginosa PCC 7806. Kljub številčnosti sistemov TA v genomu te cianobakterije jih je bilo do sedaj le nekaj tudi eksperimentalno ovrednotenih, zato smo v sklopu tega diplomskega dela želeli zapisa za dva antitoksina izraziti in izolirati iz celic Escherichia coli. Z bioinformacijskimi orodji smo zbrali in analizirali predvidene pare TA ter izmed teh izbrali dva, katerih zapisa za antitoksina smo pomnožili. Z metodami molekulskega kloniranja smo gena za antitoksina Bh695_0426 in Bh695_0430 vstavili v klonirni vektor pJET1.2/blunt. Prenos v ekspresijski vektor pET-28b(+) ni bil uspešen. Uspešno izražanje genov za antitoksina in izolacija proteinskih produktov bi omogočila nadaljnje raziskave njihove strukture in funkcije, pa tudi regulacije izražanja TA. Zaradi tega bi bilo smiselno eksperimentalno delo z manjšimi spremembami ponoviti.

Keywords

cianobakterije;Microcystis aeruginosa PCC 7806;sistemi toksin-antitoksin;antitoksini;bioinformacijska analiza;računalniška orodja;diplomska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.11 - Undergraduate Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [K. Glavič]
UDC: 575.112(043.2)
COBISS: 80181251 Link will open in a new window
Views: 306
Downloads: 71
Average score: 0 (0 votes)
Metadata: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Other data

Secondary language: English
Secondary title: Bioinformatic analysis of toxin-antitoxin pairs in the cyanobacterium Microcystis aeruginosa PCC 7806 and molecular cloning of antitoxins Bh695_0426 and Bh695_0430
Secondary abstract: Toxin-antitoxin (TA) systems are genetic elements that encode a relatively stable protein toxin that inhibits cell growth and an unstable antitoxin that inhibits the deleterious effect of the cognate toxin. Their biological function is not yet fully understood. However, they are known to participate in many biological processes, most important being postsegregational killing, abortive infection and bacterial persistence. They can be found on plasmids as well as chromosomes of numerous archaea and bacteria, including cyanobacterium Microcystis aeruginosa PPC 7806. Despite the abundance of TA systems in the genome of this cyanobacterium, only a few have been experimentally characterised so far. Therefore, our goal has been to express two antitoxins and isolate the protein products from Escherichia coli cells. Using bioinformatics tools, we collected and analysed the predicted TA pairs. From these, we selected two and amplified their antitoxin sequences. Using molecular cloning methods, we inserted the genes for the antitoxins Bh695_0426 and Bh695_0430 into the cloning vector pJET1.2/blunt. However, transfer of the antitoxins into the expression vector pET-28b(+) was unsuccessful. Successful expression of antitoxin genes and the isolation of protein products would enable further research of antitoxin function, structure and regulation of TA gene expression. It would therefore be reasonable to repeat experiments with a few adjustments.
Secondary keywords: cyanobacteria;toxin-antitoxin systems;antitoxins;
Type (COBISS): Bachelor thesis/paper
Study programme: 1000371
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, UNI Biokemija
Pages: 54 str.
ID: 13342296