Page 6 - Molecular Pathogens

Molecular Pathogens (online), 2011, Vol.2
ISSN 1925-1998 http://mp.sophiapublisher.com
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Table 1 Accession number and geographical origin and identities of CP amino acid sequences of ASPV isolates
Accession No. Isolate name Host Country Identity (%)
NC_003462 PA66 Apple Germany 71.3
AB045371 IF38 Apple Japan 71.4
DQ003336 MHczAp Apple Czech Republic 70.7
AF438521 MT32 Apple Poland 71.2
AF438522 MT24 Apple Poland 70.9
AF345892 ST54 Pear Poland 68.4
AF345893 GNKVII/34 Pear Poland 71.2
AF345894 ST132 Pear Poland 70.9
AF345895 ST113 Pear Poland 74.4
AF491929 GNKVIII/45 Pear Poland 72.9
AF491930 J335 Apple Poland 71.4
AF491931 N1 Apple Poland 70.4
AF495382 ST181 Apple Poland 71.7
AY572458 br1 Pear Brasil 71.0
EU314950 WS Apple China 71.7
EU708018 p1 Pear China 72.9
HM352726 VY1 Pear China 72.0
HM125156 PV11 Apple China 70.4
HM125157 PV2 Apple China 72.2
HM125158 PV7 Apple China 70.4
HM125159 PV5 Apple China 70.9
FJ619187 38 Apple China 71.3
FJ619188 24 Apple China 73.2
FR694922 Palampur Apple India 70.7
FN433599 Rohru Apple India 70.2
JF946772 KL9 Pear China 97.3
JF946775 KL1 Pear China 92.7
1.4 Expression and SDS-PAGE analysis
The recombinant expression plasmid ASPV-CP-Y/PET-
28a was transformed into E. col BL21 (DE3). To obtain
a highly expressed level of ASPV CP gene protein, we
tried optimizing expression conditions by using
different IPTG concentrations (0 mM, 0.5 mM, 0.8
mM, 1.0 mM, respectively) and different incubation
times (0 h, 1 h, 2 h, 3 h, 4 h, 6 h, respectively). The
results showed that the expression level of synthesized
ASPV-CP-Y/PET at 37 ℃ and with the increased
time of incubation, and the expression proteins was
increased, too. While the different concentrations of
IPTG showed insignificantly influence of expression.
A 42 kD fusion protein was highly expressed after
induced at 37℃ for 4 h with 1 mM IPTG (Figure 5).