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Volume 7, Issue 6 (Suppl)
J Biotechnol Biomater, an open access journal
ISSN: 2155-952X
World Biotechnology 2017
December 04-05, 2017
2
nd
World Biotechnology Congress
December 04-05, 2017 | Sao Paulo, Brazil
Comparative analysis of two inducible promoters for controlled nuclear transgene expression in
Chlamydomonas reinhardtii
Paula Barjona do Nascimento Coutinho, Christine Friedl Rainer Buchholz
and
Stephanie Christine Stute
Friedrich-Alexander-Universität Erlangen-Nürnberg, Germany
G
enetically, well characterized microalgae like
Chlamydomonas reinhardtii
offer the potential to photosynthetically produce high
value products such as recombinant proteins for the pharmaceutical and chemical industry. Several attempts have been made to
enhance expression of foreign genes in this green alga and in principle, allow protein production at large scale. However, satisfying
and economically attractive levels of recombinant gene products have not been achieved yet. Inducible promoters represent a useful
alternative to optimize protein yield. By providing regulated gene expression, they allow the biosynthesis of gene products at most
suitable moments of cultivation, guaranteeing higher space-time yields. In this study, two inducible promoters were compared. We
demonstrate the kinetics of induction and deactivation of the iron-responsive
Fea1
promoter and the ammonium/nitrate-responsive
Nit1
promoter in the green alga
C. reinhardtii
via the fluorescent protein mCherry and detection of mRNA levels through qPCR. Our
work lays the foundation for the establishment of a cyclic process in which promoter activity is activated and deactivated alternately
by changes in the iron and ammonium concentrations in the culture media. Fluorescence microscopy picture of
C. reinhardtii
cells
expressing mCherry under the control of the
FEA1
promoter
Biography
Paula Barjona do Nascimento Coutinho has her expertise in genetic transformation of the green alga
Chlamydomonas reinhardti
and the methods developed for the detec-
tion of the fluorescent reporter protein mCherry (flowcytometry, western blot and fluorescence microscopy).
paula.coutinho@fau.dePaula Barjona do Nascimento Coutinho et al., J Biotechnol Biomater 2017, 7:6 (Suppl)
DOI: 10.4172/2155-952X-C1-085