diplomsko delo
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: |
2021 |
Typology: |
2.11 - Undergraduate Thesis |
Organization: |
UL FKKT - Faculty of Chemistry and Chemical Technology |
Publisher: |
[K. Glavič] |
UDC: |
575.112(043.2) |
COBISS: |
80181251
|
Views: |
306 |
Downloads: |
71 |
Average score: |
0 (0 votes) |
Metadata: |
|
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 |