Molecular cloning, characterization, and immune response against white spot syndrome virus and Taura syndrome virus infections of peroxiredoxin in Litopenaeus vannamei and its antioxidant activity

in Crustaceana
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Peroxiredoxin (Prx) is an important peroxidase that can protect organisms against various oxidative stresses. In this study, a member of Prx family, designated LvPrx, was cloned from Litopenaeus vannamei. Sequence and phylogenetic analyses indicated that LvPrx belongs to the 2-Cys Prx (Prx IV) isoform. The recombinant LvPrx protein was constructed and expressed in Escherichia coli, and the purified LvPrx proteins were shown to reduce H2O2 in vitro in the presence of dithiothreitol, indicating that LvPrx is a functional peroxiredoxin. Using qRT-PCR, the mRNA expression levels of LvPrx were determined in the haemocytes of L. vannamei at different stages after being challenged with WSSV and TSV at different doses. The results showed that the expression levels of LvPrx were significantly up-regulated (P<0.05) during 4-24 h after both WSSV and TSV challenge, suggesting that LvPrx may participate in the shrimp’s immune response to viral infection.

Molecular cloning, characterization, and immune response against white spot syndrome virus and Taura syndrome virus infections of peroxiredoxin in Litopenaeus vannamei and its antioxidant activity

in Crustaceana

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References

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Figures

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    Multiple sequence alignment of Prx amino-acid sequences. The reported Prx amino-acid sequences were from GenBank and the accession numbers are as follows: Seriola lalandi (Valenciennes in Cuvier and Valenciennes, 1833) (FJ013222), Gallus gallus (Linnaeus, 1758) (XM_416800), Ixodes ricinus (Riley, 1930) (EU373821), Homo sapiens (Linnaeus, 1758) (BC016770), Penaeus monodon (Fabricius, 1798) (EU418852), and Xenopus laevis (Daudin, 1802) (JF820064).

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    Phylogenetic tree showing the relationship between LvPrx sequence with other known Prx proteins. Numbers on the branches represent bootstrap support for 2000 replicates.

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    SDS-PAGE analysis of the recombinant LvPrx protein. LvPrx was recombinantly expressed in Escherichia coli (Migula, 1895) BL21 (DE3) strain cells. Lane M, molecular weight marker; lane 1, un-induced cell lysate; lane 2, induced cell lysate; lane 3, purified LvPrx protein. This figure is published in colour in the online edition of this journal, which can be accessed via http://booksandjournals.brillonline.com/content/journals/15685403.

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    Hydrogen peroxide elimination by the recombinant LvPrx. The y-axis shows the rate of reduction of H2O2. The reactions were carried out with 10 μg recombinant Prx with 10 nM DTT or the same amount of thermo-inactivated recombinant LvPrx (iPrx) or BSA at the same condition. The error bars show standard deviation for three biological replicates. This figure is published in colour in the online edition of this journal, which can be accessed via http://booksandjournals.brillonline.com/content/journals/15685403.

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    LvPrx expression in the haemocytes of Litopenaeus vannamei (Boone, 1931) after WSSV challenge. The x-axis shows hours post virus challenge and the y-axis shows the relative expression levels of LvPrx.  Statistically significant difference (p<0.05) between the challenged group and the control group. The error bars show standard deviation for three biological replicates. This figure is published in colour in the online edition of this journal, which can be accessed via http://booksandjournals.brillonline.com/content/journals/15685403.

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    LvPrx expression in the haemocytes of Litopenaeus vannamei (Boone, 1931) after TSV challenge. The x-axis shows hours post virus challenge and the y-axis shows the relative expression levels of LvPrx.  Statistically significant difference (p<0.05) between the challenged group and the control group. The error bars show standard deviation for three biological replicates. This figure is published in colour in the online edition of this journal, which can be accessed via http://booksandjournals.brillonline.com/content/journals/15685403.

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