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Molecular & Biochemical Parasitology 148 (2006) 125132 Translational initiation in Leishmania tarentolae and
 

Summary: Molecular & Biochemical Parasitology 148 (2006) 125­132
Translational initiation in Leishmania tarentolae and
Phytomonas serpens (Kinetoplastida) is strongly
influenced by pre-ATG triplet and its 5 sequence context
Julius Lukesa,, Zdenek Parisa, Sandesh Regmia,1, Reinhard Breitlingb, Sergey Mureevc,
Susanna Kushnirc, Konstantin Pyatkovd, Milan Jirk°ua, Kirill A. Alexandrovc
a Institute of Parasitology, Czech Academy of Sciences and Faculty of Biology,
University of South Bohemia, Cesk´e Budejovice, Czech Republic
b Jena Bioscience GmbH, Jena, Germany
c Max-Planck-Institute for Molecular Physiology, Dortmund, Germany
d Division of Biology, California Institute of Technology, Pasadena, USA
Received 2 August 2005; received in revised form 13 March 2006; accepted 17 March 2006
Available online 18 April 2006
Abstract
To investigate the influence of sequence context of translation initiation codon on translation efficiency in Kinetoplastida, we constructed a
library of expression plasmids randomized in the three nucleotides prefacing ATG of a reporter gene encoding enhanced green fluorescent protein
(EGFP). All 64 possible combinations of pre-ATG triplets were individually stably integrated into the rDNA locus of Leishmania tarentolae
and the resulting cell lines were assessed for EGFP expression. The expression levels were quantified directly by measuring the fluorescence of
EGFP protein in living cells and confirmed by Western blotting. We observed a strong influence of the pre-ATG triplet on the level of protein
expression over a 20-fold range. To understand the degree of evolutionary conservation of the observed effect, we transformed Phytomonas serpens,

  

Source: Aksoy, Serap - School of Public Health, Yale University

 

Collections: Biology and Medicine