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Molecular Plant Breeding 2012, Vol.3, No.9, 91
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gel electrophoresis, mass spectrometry and bioinformatics
techniques are used to identify proteins and to record
their expression profiles, compartmentalization and
protein-protein interaction. Though, a number of
problems including detecting low abundance proteins
and capturing the dynamics of plastid proteomes are
highlighted in the literature (Baginsky and Gruissem,
2004) but recently, proteome dynamics is carried out
during plastid differentiation, using quantitative
two-dimensional gel electrophoresis and tandem
mass spectrometry in rice (Kleffmann et al., 2007).
Additionally, proteomes from stroma (Rutschow et al.,
2008; Ferro et al., 2010), thylakoid (Peltier et al.,
2000; Giacomelli et al., 2006; Rutschow et al., 2008;
Ferro et al., 2010) and envelope membranes
(Froehlich et al., 2003; Ferro et al., 2010), and from
thylakoid lumen (Peltier et al., 2002; Schubert et al.,
2002) are studied. Hence, these high-level system
biology analyses techniques are future technologies to
functionally characterize the genes, to study coordinated
expression of genes from nuclear and plastid genomes
and to study the quality traits affected by temperature
(Yun et al., 2012) during lengthy post-harvest storages.
3 Functional analyses of plastid genes using
novel approaches
In addition to characterization of plastid genes at
transcriptional and translational levels using omics
approaches, the plastid genome engineering has
facilitated the functional characterization of plastome-
encoded genes and open reading frames. Hence, the
functions of most of the major open reading frames
have now been studied through simple knockout
strategy, as reviewed by Bock and Khan (2004).
Reverse genetics, contrary to forward genes, entails
known DNA sequences that contain an open reading
frame of unknown function (s) to delete or disrupt,
resulting in lack of functional gene product with a
characterizable phenotype. The strategy involves
construction of a transformation vector in which a
selectable marker gene is cloned, between the
flanking DNA sequences,
in such a way that it either
replaces or disrupts the target gene/
ycf
from the
plastome via two events of homologous recom