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Genomics and Applied Biology 2014, Vol. 5, No. 2, 1-6
http://gab.biopublisher.ca
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these two genes both had arginase activity. We
successfully cloned these genes:
ARGAH1
,
ARGAH2
;
meantime, we got matched the sequence data on
NCBI. The full length of the cloned cDNA sequence
of
ARGAH1
is 1347 bp
；
5’ non-coding region lasts
145 bp, and the 3’ non-coding region length is 166 bp;
the ORF length is 1029 bp, encoding 342 amino acids,
and the molecular mass prediction is 37344 Da. The
predicted isoelectric point is 6.11. On the other hand,
the full length cDNA of
ARGAH2
is 1284 bp; its 5’
non-coding region lasts 34 bp, and 3’ non-coding
region is 215 bp. The ORF of
ARGAH2
is 1035 bp.
ARGAH2
encodes 344 amino acids, and the molecular
mass prediction is 37980 Da. The predicted isoelectric
point is 5.90. The similarity of
ARGAH1
and
ARGAH2
amino acids is 99%; and 86% with
Brassica
napus
, 81 % with Arginase 1 in tomato.
To further investigate the relationship between these
two genes in
Arabidopsis thaliana
, and the cognate
relation with other species, we select the arginase
genes from other species to do the comparison. The
conclusion will be drawn that the
Arabidopsis thaliana
arginase1 has the closest relationship with
Arabidopsis
lyrata
and
Brassica napus
; however, the
Arabidopsis
thaliana
arginase2 has the closest relationship with
wheat and
Brachypodium distachyon
. Two arginases
in tomato have a close relationship with each other
(Figure 1).
Figure 1 The phylogenetic tree of argianse in selected higher
plants. Species abbreviations are as follows
Lycopersicon
esculentum
(Les),
Solanum tuberosum
(Stu),
Gentiana triflora
(Gtr),
Vitis vinifera
(Vvi),
Fragaria vesca subsp.vesca
(Fve),
Malus hupehensis
(Mhu),
Ricinus communis
(Rco),
Populus
trichocarpa
(Ptr),
Cucumis sativus
(Csa),
Cicer arietinum
(Car),
Pinus taeda
(Pta),
Setaria italica
(Sit),
Prunus armeniaca
(Par),
Oryza brachyantha
(Obr),
Glycine max
(Gma),
Arabidopsis thaliana
(Ath),
Arabidopsis lyrata
(Aly),
Brassica napus
(Bna),
Medicago truncatula
(Mtr),
Brachypodium distachyon
(Bdi),
Oryza sativa
(Osa),
Triticum aestivum
(Tae)
2.2 The expression analysis of
ARGAHs
genes
during different development periods
To clarify the expression characters of arginase gene
in different growth periods in
Arabidopsis thaliana
,
this research selects dry seed, normal raised seedling
after 36 hours and 2-week seedling as experimental
material. Using QRT-PCR and SqRT-PCR methods to
research the expression of the arginase genes in different
periods of growth.
ARGAH1
and
ARGAH2
genes
expressed a lot in dry seed, and the least in 36-hour
seeding. This result shows that the expression of
arginase genes is abundant in germination, which
is consistent with the findings about these two
genes expressed a lot in tomato reproductive tissue
(Chen
et al.,
2004).
Figure 2 Expression analysis of
ARGAH1
gene and
ARGAH2
gene during different developmental stages. A represented the
expression change of
ARGAH1
gene by QRT-PCR and
SqRT-PCR; B represented the expression change of
ARGAH2
gene by QRT-PCR and SqRT-PCR