Figure legends
Figure 1. (a) Maximum likelihood phylogenetic tree displaying
the 138 PDCoV protein M sequences available in Gen Bank. In blue, the
consensus sequences used in this study. In red, 18 sequences showed
differences in branch length. (b) Comparison of 18 sequences showed
differences at amino acid level.
Figure 2 . Comparison of five models predicted by I-TASSER,
antigenic sites and surface properties. (a) Predicted models
for Consensus sequence, Chzmd2019, HKU15, M-PED CV777 and M-TGE Miller
M6. (b) Amino acid sequence for each model indicated the
predicted secondary structure, red: helix, blue: strands, black: coil.(c) Nine Antigenic sites predicted and their positions along
the consensus sequence. (d) Consensus sequence surface
properties indicating the hydrophobic and hydrophilic sites.
Figure 3 . (a) Integrity and weight verification of
PDCoV-M 654 bp synthetic gene (b) Western blot and SDS-PAGE to
verified r M-PDCoV in the expression essay. Lines 1 to 3 are an
aliquot of the culture media before cell disruption. Line 4 is an
aliquot after cell disruption. Line 5 and 6 are an aliquot after
centrifuging the cultures, soluble and insoluble phase, respectively.
The r M-PDCoV was found in insoluble phase as inclusion bodies.(c) Confirmation of r M-PDCoV after purification with
Ni-NTA agarose column with His-tag affinity. The purified recombinant
protein was observed in elutions 5 to 6.
Figure 4 . Western blot to determinate the capability ofr M-PDCoV to recognized antibodies in 17 pig blood sera from “El
Bajio” pig farm. Eight positives sera were determined and are indicate
in the image. Negative control
serum was from a sow keeping in laboratory
Figure 5. anti -PDCoV IgG distribution in sera samples
obtained from three groups of eight mice each. Black dots line indicates
PBS control group. White dots line indicates r M-PDCoV protein
group. Triangle line indicates rM-PDCoV + Iscom as an adjuvant to
enhance the immune response. Dots and triangle indicated the days of
collection sera: 7, 14, 21, and 28 Bars indicated the standard error.
The statistical significance P < 0.01 (**) and P <
0.001 (***).