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Molekulsko kloniranje in filogenetske analize nekodirajočih vtRNA
ID Nograšek, Tim (Author), ID Župunski, Vera (Mentor) More about this mentor... This link opens in a new window

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Abstract
˝Vault˝ RNA (vtRNA) je krajše, nekodirajoče zaporedje RNA, ki ga najdemo v kompleksih ˝vault˝, znotraj citoplazme evkariontskih celic. Sama vloga vtRNA zaenkrat še ni točno določena, prav tako ne vloga celotnega kompleksa. Ker se pogosto nahajajo v celicah, ki so v različnih bolezenskih stanjih, pa jih v zadnjem času vedno bolj raziskujejo. Pri človeku so bila identificirana štiri različna zaporedja vtRNA in sicer vtRNA1-1, vtRNA1-2, vtRNA1-3 in vtRNA2-1. Slednji se nahaja na drugem lokusu petega kromosoma kot ostali trije. Poleg teh štirih zaporedij sta v človeškem genomu prisotna dva psevdogena, poimenovana vtRNA2-2 in vtRNA3-1. Ta dva se nahajata na drugih kromosomih kot ostali štirje. V okviru diplomske naloge smo naredili filogenetsko analizo zaporedij vtRNA pri vretenčarjih. Ugotovili smo, da se razvoj različnih vrst vtRNA delno ujema s splošnim taksonomskim razvojem, hkrati pa je prisotnih tudi nekaj odstopanj. Pri nekaterih odstopanjih smo z podrobnim pregledom poravnav zaporedij pokazali njihov razlog. Največja ujemanja zaporedij vtRNA so bila na regulacijskih mestih, kjer so se ta skozi evolucijo najbolj ohranila. Pri laboratorijskem delu pa smo pripravili vektorje z različnimi človeškimi zaporedji vtRNA, da bi lahko izražali v sesalskih celicah. Na ta način bomo lahko določili njihove interakcijske partnerje in lokacijo v celici.

Language:Slovenian
Keywords:vtRNA, filogenetska analiza, kloniranje IVA
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2021
PID:20.500.12556/RUL-130006 This link opens in a new window
COBISS.SI-ID:82872579 This link opens in a new window
Publication date in RUL:10.09.2021
Views:1155
Downloads:187
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Secondary language

Language:English
Title:Molecular cloning and phylogenetic analyses of noncoding vtRNA
Abstract:
Vault RNA (vtRNA) is a shorter, noncoding RNA sequence found in vault complexes in the cytoplasm of eukaryotic cells. The function of vtRNA is still not known, nor is the function of the complex, but because they are found in various unusual states of cells, there has been increased research recently. Four different vtRNA sequences have been identified in the human genome: vtRNA1-1, vtRNA1-2, vtRNA1-3 and vtRNA2-1. The last one is also the one found at different locus of chromosome number five than the other three. In addition to these four RNA molecules, there are also two pseudogenes in the human genome: vtRNA2-2 and vtRNA3-1. These two are located on different chromosomes than the other four vtRNAs. We performed a phylogenetic analysis of vtRNAs in vertebrates. We found that the development of different types of vtRNAs is consistent with the general taxonomic evolution, but at the same time there are some divergences. For some divergences we were able to explain them with a detailed review of sequence alignments, while for others the reason for their placement in the phylogenetic tree was different point mutations. In the laboratory part, we made vectors with different types of vtRNAs that can be expressed in mammalian cells. In this way, we enable further research on their interaction partners and cellular location.

Keywords:vtRNA, phylogenetic analysis, IVA cloning

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