Kloniranje i ekspresija ljudskog proteina BACH2 u bakterijskim i animalnim stanicama

master's thesis

• Title (english): Cloning and expression of human BACH2 protein in bacterial and animal cells
• Author: Nina Ivanković
• Mentor: Vlatka Zoldoš (mentor)
• Committee member: Vlatka Zoldoš (predsjednik povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Damjan Franjević (član povjerenstva)
• Committee member: Ivana Ivančić Baće (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2017-06-28, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Ljudski protein BACH2 transkripcijski je faktor specifično eksprimiran u makrofagima, limfocitima T i B. U limfocitima B, BACH2 regulira ranu diferencijaciju limfocita B iz zajedničkog multipotentnog progenitora i terminalnu diferencijaciju plazma stanice iz zrelog limfocita B. Cjelogenomske asocijacijske studije (GWAS) povezale su gen BACH2 s N-glikozilacijom imunoglobulina G (IgG) te s nekim upalnim bolestima poput upalnih bolesti crijeva, reumatodnog artritisa i sistemskog lupusa eritematodesa. Alternativna glikozilacija antitijela IgG određuje njegovu funkciju – dodatak galaktoze i sijalinske kiseline na glikansku strukturu daje antiupalno djelovanje ovom antitijelu. Gen BACH2 također je uključen u imunu homeostazu i imunosenescenciju, a poznato je da tijekom imunosenescencije antitijelo IgG postaje proupalno. Polimorfizam jednog nukleotida (SNP) u genu BACH2 povezan je s galaktozilacijom glikana na antitijelu IgG, što čini ovu molekulu proupalnom. Stoga su potrebne detaljne funkcionalne studije koje bi utvrdile točnu ulogu gena BACH2 u glikozilaciji IgG-a povezanu s njegovom funkcijom u upalnim bolestima te u imunosenescenciji. Cilj istraživanja provedenih u sklopu ovog diplomskog rada bilo je klonirati i eksprimirati protein BACH2 u bakterijskim stanicama Rosetta te prekomjerno eksprimirati ovaj protein u animalnim stanicama HEK293T.

Abstract (english)

Human protein BACH2 is a transcription factor specifically expressed in macrophages and T and B lymphocytes. In B lymphocytes, BACH2 regulates early differentiation of B lymphocyte from common multipotent progenitor and terminal differentiation of plasma cells from naive B lymphocyte. Genome-wide association studies (GWAS) associated BACH2 gene with N-glycosylation of immunoglobulin G (IgG), as well as with some inflammatory diseases such as inflammatory bowel diseases, rheumatoid arthritis and systemic lupus erithematosus. Alternative glycosylation of IgG defines its function – addition of galactose and sialic acid on glycan structure regulates antiinflammatory function of the antibody. The BACH2 gene is shown to be implicated in immune homeostasis and immunosenescence, during which IgG becomes proinflammatory. Single nucleotide polymorphisms (SNP) in BACH2 gene have been associated with galactosylation of glycans on IgG which makes this molecule proinflammatory. To unravel the exact role of BACH2 gene in glycosylation of IgG associated with its function in inflammatory diseases and immunosenescence, more detailed functional studies are needed. The goal of the investigations conducted within this graduation thesis was to clone and to express BACH2 protein in bacterial cells Rosetta and to overexpress the same protein in animal HEK293T cells.

• Keywords: BACH2; transkripcija; glikani; upalne bolesti
• Keywords (english): BACH2; transcription; glycans; inflammatory disease
• Language: croatian
• URN:NBN: urn:nbn:hr:217:317542
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-09-07 10:06:09