diplomsko delo
Maja Trifkovič (Avtor), Vera Župunski (Mentor)

Povzetek

Nekodirajoče RNA Y so bile odkrite pri večini vretenčarjev, njihovi homologi ali njim podobne molekule pa tudi pri drugih organizmih. Imajo značilno sekundarno strukturo, sestavljeno iz stebel in zank in se povezujejo s številnimi proteini, najpogosteje s proteinoma Ro60 in La. Čeprav vse njihove funkcije še niso znane, naj bi sodelovale pri iniciaciji replikacije kromosomske DNA in v kompleksu z Ro60 pri vezavi in razgradnji nepravilno zvitih RNA. V okviru eksperimentalnega dela diplomske naloge smo z metodo kloniranja s sestavljanjem in vivo (kloniranje IVA) uspešno vstavili zapise za vse štiri človeške RNA Y (hY1, hY3, hY4 in hY5) v vektor pcDNA3.1-U6 za njihovo izražanje v sesalskih celicah. Drugi del diplomske naloge pa predstavljajo filogenetske analize zaporedij RNA Y iz različnih organizmov in primerjava s sorodstvenimi odnosi znotraj sesalcev. Ugotovili smo, da so zaporedja RNY1 in RNY3 iz različnih organizmov zelo podobna, medtem ko so zaporedja RNY4 in RNY5 bolj različna. S filogenetsko analizo človeških RNA Y smo ugotovili, da so te najverjetneje nastale s podvojitvijo genov. Najprej je prišlo do podvojitve gena za hY4, nastal je gen za hY1, ki se je prav tako podvojil in iz njega sta nastala gena za hY3 in hY5. Našli smo tudi šest različnih zaporedij RNA Y opice rezus makak (Macaca mulatta), pri kateri je najverjetneje prišlo še dodatnih podvojevanj genov. S filogenetsko analizo genov vsake od štirih RNA Y iz različnih organizmov smo ugotovili, da filogenija RNA Y ne ustreza popolnoma sorodstvenim odnosom znotraj sesalcev. Zaporedja RNA Y mogoče sicer niso najbolj primerna izbira za tovrstne analize, saj so kratka in med seboj zelo podobna. Prav tako je bil nabor zaporedij iz različnih organizmov neuravnotežen (prevladovala so zaporedja primatov, ki so bila večinoma identična).

Ključne besede

nekodirajoče RNA Y;kloniranje IVA;filogenetska analiza;diplomska dela;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.11 - Diplomsko delo
Organizacija: UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik: [M. Trifkovič]
UDK: 577.21(043.2)
COBISS: 130880515 Povezava se bo odprla v novem oknu
Št. ogledov: 49
Št. prenosov: 14
Ocena: 0 (0 glasov)
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Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Molecular cloning and phylogenetic analyses of noncoding Y RNAs
Sekundarni povzetek: Noncoding Y RNAs have been discovered in most vertebrates, and their homologs or similar molecules have also been discovered in other organisms. They have a typical secondary structure consisting of stems and loops and interact with many different proteins, mainly Ro60 and La. Although not all of their functions are known, they have been shown to play a role in the initiation of chromosomal DNA replication and, in a complex with Ro60, in the binding and degradation of misfolded RNAs. In the experimental part of the thesis, we incorporated the genes for all four human Y RNAs (hY1, hY3, hY4 and hY5) into the vector pcDNA3.1-U6 for expression in mammalian cells by in vivo assembly cloning (IVA cloning). The second part of the thesis is phylogenetic analyses of Y RNAs from different organisms and comparisons with mammalian relationships. We found that the sequences of RNY1 and RNY3 from different organisms are very similar, whereas the sequences of RNY4 and RNY5 are more different. In the phylogenetic analysis of human Y RNAs, we found that they most likely arose by gene duplications. First, the hY4 gene was duplicated to give rise to hY1, which also duplicated to give rise to the genes for hY3 and hY5. We found six different Y RNA sequences from rhesus macaque (Macaca mulatta) in which further gene duplication most likely occurred. Through phylogenetic analysis of the four Y RNA genes from different organisms, we found that the phylogeny of Y RNAs is not completely consistent with mammalian relationships. The sequences of Y RNAs were perhaps not the most appropriate choice for such analyses because they are short and very similar to each other. In addition, we had a very unbalanced set of sequences from different organisms (most of them were primate sequences that were mostly identical).
Sekundarne ključne besede: Y RNA;IVA cloning;phylogenetic analysis;RNK;Univerzitetna in visokošolska dela;
Vrsta dela (COBISS): Diplomsko delo/naloga
Študijski program: 1000371
Konec prepovedi (OpenAIRE): 1970-01-01
Komentar na gradivo: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, UNI Biokemija
Strani: 74 str
ID: 16479228