master's thesis
Vector construction for heterologous expression At4g15590 gene product from plant A. thaliana(L.) Heynh

Krešimir Viandt (2012)
Sveučilište Josipa Jurja Strossmayera u Osijeku
Odjel za biologiju
Metadata
TitleKonstrukcija vektora za hetrolognu ekspresiju produkta gena At4g15590 iz biljke A. thaliana(L.) Heynh
AuthorKrešimir Viandt
Mentor(s)Hrvoje Fulgosi (thesis advisor)
Abstract
U genomu biljke A. thaliana L. Heynh, identificiran je gen At4g15590 koji kodira suppressor of white apricot (SWAP) regulator prekrajanja (engl. splicing). Mikrosekvencioniranjem AM8 frakcijeizolirane iz cytb6f kompleksa iz špinata (S. oleracea) utvrđen je slijed PELDSSALV, koji pokazuje veliku homologiju sa slijedom PELDESALV u biljci A. thaliana (SWAP protein). Gen veličine 1677pb, koji je sastavljen je od 4 ili 5 egzona ovisno o višestrukim varijantama transkripata, na aminoterminalnom kraju gena karakterizira surp-domena, a na karboksiterminalnom kraju ubikvitin-domena.Metodom lančane reakcijom polimerazom (PCR) umnožen je dio gena SWAP, veličine 350 nukleotida, i ugrađen u plazmidni vektor pRSETa, gdje je fuziran na N-terminalnom kraju s peptidnim biljegom (6xHis tag). Ekspresija proteina ispitivana je u različitim bakterijskim stanicama pri različitim uvijetima uzgoja.
KeywordsSWAP splicing pRSETa vector lac operon protein expression
Parallel title (English)Vector construction for heterologous expression At4g15590 gene product from plant A. thaliana(L.) Heynh
Committee MembersVera Cesar (committee chairperson)
Hrvoje Fulgosi (committee member)
Elizabeta Has Schon (committee member)
GranterSveučilište Josipa Jurja Strossmayera u Osijeku
Odjel za biologiju
PlaceOsijek
StateCroatia
Scientific field, discipline, subdisciplineNATURAL SCIENCES
Biology
Study programme typeuniversity
Study levelgraduate
Study programmeMaster of Science Graduate University Study Programme in Biology; specializations in: Master of Biology
Study specializationMaster of Biology
Academic title abbreviationmag.biol.
Genremaster's thesis
Language Croatian
Defense date2012-09-26
Parallel abstract (English)
In plants A. thaliana L. Heynh genome a gene At4g15590 that codes suppressor of white apricot(SWAP) regulator for splicing has been discovered. Via microsequencing of AM8 fraction isolated from cytb6f spinach complex (S. oleracea) a sequence PELDSSALV has been established, a sequence that shows great hegemony with PELDESALV sequence in A. thaliana plant (SWAP protein). A 1677pb sized-gene, constructed from 4 or 5 exons, depending on multiple transcript variants, is characterized by a surp-domaine at the aminothermal gene region, and ubiquitin-domain at the carboxy-terminal region. Via polymerase chain reaction (PCR), SWAP part of the gene, 350 nucleotides in size, has been multiplied and built in a plasmidic pRSET vector where it has been fused on N-terminal end with peptide (6xHis) tag. Protein expresion has been researched in various bacterial cells under different bacteria breeding conditions.
Parallel keywords (Croatian)SWAP prekrajanje vektor pRSETa lac operon ekspresija proteina
Resource typetext
Access conditionOpen access
Terms of usehttp://rightsstatements.org/vocab/InC/1.0/
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:181:727613
CommitterVesna Radman Meic