No public access
master's thesis
Heterologous expression and purification of recombinant protein TROL for the purpose of antiserum production in rabbits

Alma Salim (2014)
Sveučilište Josipa Jurja Strossmayera u Osijeku
Odjel za biologiju
Metadata
TitleHeterologna ekspresija i pročišćavanje rekombinantnog proteina TROL u svrhu proizvodnje antiseruma u zečevima
AuthorAlma Salim
Mentor(s)Hrvoje Fulgosi (thesis advisor)
Abstract
Regulatorni gen At4g01050 smješten na 4. kromosomu biljke Arabidopsis thaliana (L.) Heynh kodira za protein TROL (thylakoid rhodanase-like protein). TROL je integralni membranski protein potreban za vezanje FNR-a (feredoksin-NADP+ reduktaze) i održavanje visoke efikasnosti linearnog toka elektrona u fotosintezi. Heterologna ekspresija dopušta proizvodnju biljnih proteina u organizmu koji je različit od prirodnog izvora. Za povećanu ekspresiju proteina TROL odabran je ekspresijski vektor pRSET A, u kojem je uklonirani gen pod kontrolom T7 promotora. Soj E. coli BL21(DE3)pMico transformiran je ekspresijskim vektorom pRSET A/miniTROL te je ekspresija proteina inducirana dodatkom IPTG-a. Dobiveni rekombinantni protein je zatim pročišćen afinitetnom kromatografijom u nativnim uvjetima i korišten kao antigen za proizvodnju antiseruma u zečevima. Ovako dobiveni antiserum obično je specifičniji i imunoreaktivniji nego antitijela razvijena na kratke sintetske peptide
KeywordsTROL recombinant protein protein expression protein purification antiserum
Parallel title (English)Heterologous expression and purification of recombinant protein TROL for the purpose of antiserum production in rabbits
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 date2014-11-28
Parallel abstract (English)
The regulatory gene At4g01050 is located on the 4th chromosome of Arabidopsis thaliana (L.) Heynh, encoding for protein TROL (thylakoid rhodanase-like protein). TROL is an integral membrane protein required for tethering of FNR (ferredoxin-NADP+ reductase) and sustaining efficient linear electron flow in vascular plants. Heterologous expression allows the production of plant proteins in an organism which is simpler than the natural source. For the increased expression of TROL protein, pRSET A vector has been selected, into which our gene of interest is cloned under the control of the T7 promotor. E. coli strain BL21(DE3)pMico was transformed with the pRSET A/miniTROL construct and expression was induced by the addition of IPTG. The resulting recombinant protein was then purified by affinity column chromatography under native conditions and has been used as an antigene for antiserum production in rabbits. The resulting antiserum is expected to be more specific and immunoreactive than the antibodies raised against shorter synthetic peptides.
Parallel keywords (Croatian)TROL rekombinantni protein ekspresija proteina pročišćavanje proteina antiserum
Resource typetext
Access conditionNo public access
Terms of usehttp://rightsstatements.org/vocab/InC/1.0/
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:181:477600
CommitterVesna Radman Meic