Evolutionary analysis of MGgLYZ1 and MGgLYZ2 genes.

<p>Evolutionary analysis of MGgLYZ1 and MGgLYZ2 genes.</p

Effect of pH, temperature and lysozyme inhibitor (PliG) on the activities of MGgLYZ1 (open symbols) and MGgLYZ2 (filled symbols).

<p>Lysozyme activity is shown as % of the highest activity. The values were shown as mean ± SD (n = 3).</p

Analysis of recombinant MGgLYZ1 and MGgLYZ2 proteins and antibodies.

<p>(a) SDS-PAGE analysis, LC-ESI-MS/MS identification of MGgLYZ1 (b) and MGgLYZ2 (c), western blotting analysis of MGgLYZ1 (d) and MGgLYZ2 (e) polyclonal antibodies.</p

The sampling sites of <i>Mytilus galloprovincialis</i> along the coastal area of northern China.

<p>The sampling sites of <i>Mytilus galloprovincialis</i> along the coastal area of northern China.</p

Multiple alignments of MGgLYZ1 and MGgLYZ2 with other mollusk gLYZs deposited in GenBank.

<p>The black shadow region indicates positions where all sequences share the same amino acid residue. Gaps are indicated by dashes to improve the alignment. The asterisk indicates the conserved cysteine residues and the pound sign indicates the conserved active residues in mollusk gLYZs. The GenBank accession no. and the species are as follows: JQ244770 (<i>Mytilus galloprovincialis</i> 1, Mg1), JQ244771 (<i>Mytilus galloprovincialis</i> 2, Mg2), DQ227696.1 (<i>Chlamys farreri</i>, Cf), AY788903 (<i>Argopecten irradians</i>, Ai), GR867752.1 (<i>Mizuhopecten yessoensis</i>, My), ES392226.1 (<i>Mytilus californianus</i>, Mc), FC670738.1 (<i>Lottia gigantean</i>, Lg), DC603639.1 (<i>Nesiohelix samarangae</i>, Ns), ES491677.1 (<i>Biomphalaria glabrata</i>, Bg), EV289297.1 (<i>Tritonia diomedea</i>, Td), ADV36303.1 (<i>Physella acuta</i>, Pa), FK716269.1 (<i>Ilyanassa obsolete</i>, Io), GW425811 (<i>Oncomelania hupensis</i> 1, Oh1), GW426148 (<i>Oncomelania hupensis</i> 2, Oh2) and GW427036 (<i>Oncomelania hupensis</i> 3, Oh3).</p

Phylogenetic tree constructed by neighbour-joining method based on the sequences of gLYZs from different animals.

<p>Numbers at the forks indicate the bootstrap values (in %) out of 1000 replicates. The sequences used to construct phylogeny trees of gLYZs are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045148#pone.0045148.s005" target="_blank">Table S3</a>.</p

Tissue-specific expression of MGgLYZ1 (a) and MGgLYZ2 (b) mRNA.

<p>The mRNA expression level is calculated relative to actin expression and shown as mean ± SD (n = 4). Significant difference is indicated with an asterisk at <i>P</i><0.05.</p

The genomic structures of MGgLYZ1 and MGgLYZ2 compared with those of Japanese flounder and chicken gLYZ genes.

<p>Black boxes indicate the corresponding open reading frames in the mussel, flounder and chicken genes. White boxes indicate the untranslated regions. The size of each intron is shown near each intron region.</p

Predicted three-dimensional structure of MGgLYZ1 and MGgLYZ2.

<p>a: Three-dimensional structure predicted by SWISSMODEL. b: Electrostatic surface potentials of MGgLYZ1 and MGgLYZ2. The color runs from intense red (lowest) to intense blue (highest) potential.</p

Temporal expression profiles of gLYZ mRNAs in hemocytes (a-MGgLYZ1, b-MGgLYZ2) and hepatopancreas (c-MGgLYZ1, d-MGgLYZ2) after bacterial challenge.

<p>The mRNA expression level is calculated relative to actin expression and shown as mean ± SD (n = 4). Significant difference from control (0 h) is indicated with an asterisk at <i>P</i><0.05.</p