Production of interferon α2b from green microalga Chlamydomonas reinhardtii

Postgraduate Thesis uoadl:1319320 145 Read counter

Unit:
ΠΜΣ Μικροβιακή Βιοτεχνολογία
Library of the School of Science
Deposit date:
2015-11-19
Year:
2015
Author:
Ψυχάρη Ελένη
Supervisors info:
Γιώργος Διαλλινάς Καθηγητής
Original Title:
Παραγωγή ιντερφερόνης α2b από το σύστημα του χλωροφύκους Chlamydomonas reinhardtii
Languages:
Greek
Translated title:
Production of interferon α2b from green microalga Chlamydomonas reinhardtii
Summary:
Human interferon IFNα2b is a widely used antivirus drug, which is nowdays
produced by Escherichia coli. As a bacterium, E.coli is unable to glycosylate
interferon. In order to have the requisite halflife in patient’s blood,
interferon is
glycosylated in vitro (addition of poly-ethyleno glycol). There have been made
lots of
efforts to express interferon from other recombinant expression systems, however
with no success for industrial production.
Microalgae are a diverse group of photosynthetic microorganisms with significant
biotechnological possibilities, which make them attractive expression systems
for
pharmaceutical and biotechnological proteins. The green microalga Chlamydomonas
reinhardtii has been served as a model organism for over 60 years, and thus it
offers
the most prevalent molecular toolkit among the microalgae. Some of its
advantages
are its speed growth, ease to handle, ability to scale up, low cultivation
cost, and the
fact that it is regarded as safe (GRAS). Moreover, the expression of recombinant
proteins from the nucleus of C.reinhardtii offers the advantages of eukaryotic
expression systems, such as inducible transgene expression, correct folding of
complex proteins, subcellular location and secretion of recombinant proteins,
and
post-translational modifications.
The present study is an effort to express human IFNα2b from the nucleus of
C.reinhardtii. The optimized gene of human IFNα2b for C. reinhardtii was cloned
into pChlamy_1 plasmid, with whom wild type cells of C.reinhardtii were
transformed with, by electroporation. The transformed colonies that occurred by
antibiotic resistance to hygromycin B, were checked for the insertion of the
ifn gene
by colony PCR. Subsequently, the mRNA of the positive colonies was extracted and
the expression of the ifn gene was detected by real-time PCR. The results of
this study
were promising for further investigation of this expression system.
Keywords:
Interferon α2b, Chlamydomonas reinhardtii, Expression, Microalgae transformation, Microalgae
Index:
No
Number of index pages:
0
Contains images:
Yes
Number of references:
86
Number of pages:
90
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