INVESTIGADORES
PETRILLO Ezequiel
congresos y reuniones científicas
Título:
PROTOPLASTS AS A TOOL TO STUDY ALTERNATIVE SPLICING IN ARABIDOPSIS
Autor/es:
MICAELA A. GODOY HERZ; EZEQUIEL PETRILLO; ALBERTO R. KORNBLIHTT
Reunión:
Congreso; SAIB - XLVIII Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2012
Resumen:
Arabidopsis protoplasts are cells without cell walls obtained from
leaves after an enzymatic treatment. Protoplasts maintain many of
the physiological properties of the plants which they come from.
We have established the experimental conditions to obtain
Arabidopsis mesophyl protoplasts in order to asess alternative
splicing responses to light/darkness treatments as previously
observed in Arabidopsis seedlings. We use the RS31
transcriptional unit as a model. In seedlings, light treatment causes
an increase of the functional mRNA isoform. Protoplasts seem to
show a light/darkness effect, only slightly lower than in seedlings.
Furthermore, we have constructed a series of alternative splicing
reporter minigenes to be introduced into the protoplasts through
PEG-mediated transformation. These minigenes combine different
alternative splicing events ?exon cassette and intron retention or
alternative 5? splice sites- with three different promoters. The idea
is to evaluate if promoters affect alternative splicing in protoplasts
as it was described in animal cells. Our protoplast transformation
controls using a GFP reporter include detection of the expressed
protein by epifluorescence microscopy and of expressed RNA by
RT-PCR. Our results indicate that protoplast isolation and
transformation are a useful new tool to study alternative splicing in
plants.