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Bioscience Methods 2014, Vol.6, No.1, 1-13
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3
identity between the cDNA ORF and the
StOSM
-3B
ORF. An alignment with the amino acid sequence
showed that the putative amino acid sequences
encoded by the isolated cDNA ORF have six residual
substitutions compared with the sequence of
StOSM
-3B (Figure 2).
Figure 2 Alignment of the cDNA ORF isolated with
StOSM
-3B by putative amino acid sequence. The sequence alignment was
performed using the ClustalW2 online software in EMBL-EBI. The green colour indicates the mutated amino acids (
StOSM
-3Bm:
St
OSM-3B isolated in this study;
StOSM
-3Bc:
StOSM
-3B isolated by Castillo RA et al [10])
The isolated cDNA in this study was still named after
StOSM
-3B because the cDNA sequence showed high
identity with that of
StOSM
-3B. Analysis of the
expression of
StOSM
-3B in
E. coli
indicated that as
the
StOSM
-3B mRNA accumulates, the tolerance of
recombinant
E. coli
, as described by colony livability
under osmotic stress, was significantly improved,
even as the osmotic stress generated by PEG6000
increased (Hu et al 2012).
Wounding, abscisic acid (ABA), low temperature,
NaCl and salicylic acid (SA) increased the
StOSM
-3B (pA81) mRNA expression in
S.
commersonii
cell cultures and in plants grown
in
vitro
. The reason for why these conditions induce
StOSM
-3B mRNA accumulation is unknown yet
(Zhu et al. 1995). In addition, the involvement of
StOSM
-3B in water stress has not yet been reported.
1.1.2
StOSM
genes in potato
The
sequence of the cloned
StOSM
-3B cDNA was
retrieved in the PGSC_DM_v3.4_gene.fasta file from the
Potato Genome Sequencing Consortium website
(http://solanaceae.plantbiology.msu.edu) (Cory et al.
2014). A total of eleven
StOSM
genes distributed among
five chromosomes in the potato genome were identified
from the PGSC database. The sequence structure of the
eleven potato
StOSM
cDNAs is shown in Table 1.
All potato
StOSM
genes were collected from the
NCBI GenBank using BLAST analysis and aligned
with these eleven
StOSM
. No one beyond eleven
StOSM
was found according the sequence alignment.
Therefore, seven genes, such as
StOSM
-3B and -3F,
were designated according to names used previously
and registered in the database (Castillo et al. 2005).
Four new
StOSM
genes were identified and named
based on the length of the putative encoded peptide.