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Molecular Pathogens (online), 2011, Vol.2
ISSN 1925-1998 http://mp.sophiapublisher.com
36
Figure 3 Phylogenetic tree constructed from the complete CP
gene at aa sequences of 28 ASPV isolates
Figure 4 Identification of recombined plasmid by PCR and
Restriction enzyme
Note: M: D514A 200 marker; M1: SM1243 marker; 1~3:
Identification by PCR; 4: Recombined plasmid; 5: Identification of
recombined plasmid by double restriction enzyme
Figure 5 SDS-PAG of expression products of ASPV CP gene
Note: M: D523A marker; 1: Vector control; 2: Total proteins
from BL/ASPV-CP-Y/PET-28a uninduced by IPTG; 3~7: Total
proteins from BL/ASPV-CP-Y/PET-28a induced by IPTG for 1 h,
2 h, 3 h, 4 h, 6 h, respectively
2 Discussion
In this research, we reported the complete genomic
sequences of Chinese ASPV isolates Y, and the
complete CP gene consisted of 1 194 nucleotides and
encoded a polypeptide of 397 amino acid (aa), which
has been deposited in the GenBank and allocated the
accession numbers of JF946774. Comparison of the
amino acid sequences of Ya pear CP gene with other
ASPV isolates showed approximately 70% similarity.
Sequencing and analysis of ASPV isolates revealed
that the virus had complex sequence variants among
different isolates. Figure 3 showed that ASPV isolates
at aa sequence demonstrated three groups. All ASPV
isolates from apple were clustered to group I, whereas
those of pear were clustered to groups
Ⅱ
(except
NC_003462) and the Ya pear were clustered to group
Ⅲ
. The isolates in group I and group
Ⅱ
mostly were
from Poland and China, and the two groups showed
that there were no clear geographical differentiations
between ASPV isolates and slight related to host, and
this result was similar to Li et al (2010a). Furthermore,
group I and group II both can be further divided into
five sub-groups. However, the formation of these
sub-groups had not certain regularity. According to
the classification of the ASPV CP gene, we suggested
that the variation of ASPV isolates were closely
related to the host and ecological environment.
Therefore, it is necessary to consider the geography,
climates, as well as other part of sequences
information of genomic to analyze the variation and
evolutionary relationships of ASPV.