Biology Reference
In-Depth Information
DNA and protein metabolism, cell wall biogenesis, cytoskeleton
and, cellular cycle and translocation. The percentage of total pro-
teins identifi ed which were present in one fraction was: 25, 23, 15,
and 23 % for the albumin, globulin, prolamin, and glutelin frac-
tions, respectively. We have established the fi rst
Q. ilex
acorn refer-
ence proteome that can be used in subsequent analysis, namely, the
identifi cation of protein markers to discriminate between popula-
tions, studies on the germination process, etc.
The main limitation of this approach is that the genus
Quercus
is an orphan plant species poorly characterized at the genetic and
physiological levels. The low number of well annotated genomic
sequences available for genus
Quercus
, and for
Q. ilex
in particular,
in public databases limits the confi dent protein identifi cation.
Moreover, most software tools have been developed to search in a
non-error tolerant way against a database of known proteins [
10
,
23
].
A higher number of identifi cations which are more accurate would
be obtained by searching against EST sequences [
25
,
26
]. These
sequences can be obtained from the following EST public database
(
http://compbio.dfci.harvard.edu/tgi/
;
http://www.fagaceae.
group is constructing an EST database for
Quercus.
4
Notes
1. References to specifi c companies, equipment or pieces of
equipment are not mandatory and do not represent an endorse-
ment by the authors
2. Standard reagents and items of current use, such as Bovine
serum albumin (BSA), liquid nitrogen and polyvinylpolypyr-
rolidone, are not specifi ed.
3. Protein extraction and fractionation procedures were based on
the differential solubility criteria described by Osborne (1924),
and later adapted to
Quercus
subsp. by Fonseca et al. (1997)
and Martin et al. (2009) [
2
,
27
,
28
].
4. The extraction is performed with 10 g of fresh tissue because
fractions of albumins, globulins and prolamins are minority
proteins, with 1 g of tissue the small amount of protein iso-
lated in the mentioned fraction is not enough to analyse.
5. This step is to reduce lipid content, which allows the obtaining
of a higher quality extract for subsequent analysis.
6. Be careful that the pellet does not stick tightly to the bottom
of the tube.
7. This step is to remove traces of diethylether.
