“…These primer sets were used to amplify the corresponding PCR products as templates for dsRNA synthesis. The dsRNA was synthesized using T7 polymerase (Fermentas, USA) following the previous method (Wang et al, 2009).…”
Section: Rna Interference Assaymentioning
confidence: 99%
“…In Drosophila S2 cells (Schneider line 2), Ran protein, a small Ras-like GTPase, was involved in the regulation of phagocytosis against Drosophila C virus (DCV). Some members of the ADP ribosylation factors (Arfs) found in mammals (D'Souza-Schorey and Chavrier, 2006;Nie et al, 2003) and the shrimp Marsupenaeus japonicas Wang et al, 2009;Zhang et al, 2010), which are classified with small GTP-binding proteins (~21 kDa), are involved in virus infection. The relationship between various Arfs and virus invasion is thus worth investigating.…”
ADP-ribosylation factors (Arfs) are small GTP-binding proteins that have an essential function in intracellular trafficking and organelle structure. To date, little information is available on the Arfs in the economically important giant freshwater prawn Macrobrachium rosenbergii and their relationship to viral infection. Here we identified two Arf genes from M. rosenbergii (MrArf1 and MrArf2) for the first time. Phylogenetic analysis showed that MrArf1, together with MjArf1 from shrimp Marsupenaeus japonicus belonged to Class I Arfs. By contrast, MrArf2 didn't not match any of the Arfs classes of I/II/III, although it could be clustered with an Arf protein from M. japonicas called MjArfn, which may represent an analog of the Arf. MrArf1 was ubiquitously expressed in all the examined tissues, with the highest transcription level in the hepatopancreas, whereas MrArf2 was only highly expressed in the hepatopancreas and exhibited very low levels in the heart, stomach, gills and intestine. The expression level of MrArf1 in the gills was down-regulated post 24 h WSSV challenge, and reached the maximal level at 48 h. MrArf1 in the hepatopancreas went up from 24 to 48 h WSSV challenge. MrArf2 transcript in the gill also went down at 24 h and then was upregulated at 48 h WSSV challenge. MrArf2 increased significantly in the hepatopancreas 24 h after infection and then went down at 48 h WSSV challenge. RNAi results showed that knockdown of MrArf1 or MrArf2 could inhibit the expression of the envelope protein gene vp28 of the WSSV. So, it could be speculated that MrArf1 and MrArf2 might play important roles in the innate immune system against WSSV infection.
“…These primer sets were used to amplify the corresponding PCR products as templates for dsRNA synthesis. The dsRNA was synthesized using T7 polymerase (Fermentas, USA) following the previous method (Wang et al, 2009).…”
Section: Rna Interference Assaymentioning
confidence: 99%
“…In Drosophila S2 cells (Schneider line 2), Ran protein, a small Ras-like GTPase, was involved in the regulation of phagocytosis against Drosophila C virus (DCV). Some members of the ADP ribosylation factors (Arfs) found in mammals (D'Souza-Schorey and Chavrier, 2006;Nie et al, 2003) and the shrimp Marsupenaeus japonicas Wang et al, 2009;Zhang et al, 2010), which are classified with small GTP-binding proteins (~21 kDa), are involved in virus infection. The relationship between various Arfs and virus invasion is thus worth investigating.…”
ADP-ribosylation factors (Arfs) are small GTP-binding proteins that have an essential function in intracellular trafficking and organelle structure. To date, little information is available on the Arfs in the economically important giant freshwater prawn Macrobrachium rosenbergii and their relationship to viral infection. Here we identified two Arf genes from M. rosenbergii (MrArf1 and MrArf2) for the first time. Phylogenetic analysis showed that MrArf1, together with MjArf1 from shrimp Marsupenaeus japonicus belonged to Class I Arfs. By contrast, MrArf2 didn't not match any of the Arfs classes of I/II/III, although it could be clustered with an Arf protein from M. japonicas called MjArfn, which may represent an analog of the Arf. MrArf1 was ubiquitously expressed in all the examined tissues, with the highest transcription level in the hepatopancreas, whereas MrArf2 was only highly expressed in the hepatopancreas and exhibited very low levels in the heart, stomach, gills and intestine. The expression level of MrArf1 in the gills was down-regulated post 24 h WSSV challenge, and reached the maximal level at 48 h. MrArf1 in the hepatopancreas went up from 24 to 48 h WSSV challenge. MrArf2 transcript in the gill also went down at 24 h and then was upregulated at 48 h WSSV challenge. MrArf2 increased significantly in the hepatopancreas 24 h after infection and then went down at 48 h WSSV challenge. RNAi results showed that knockdown of MrArf1 or MrArf2 could inhibit the expression of the envelope protein gene vp28 of the WSSV. So, it could be speculated that MrArf1 and MrArf2 might play important roles in the innate immune system against WSSV infection.
“…e l s e v i e r . c om / l oc a t e / f s i challenge [13] suggesting that shrimp Arfs may also respond to virus infection. Even though a number of Arfs have been isolated and characterized from microorganisms as well as mammalian cells, there are still few documents about Arfs from shrimp to date.…”
Section: Contents Lists Available At Sciencedirectmentioning
“…Total RNAs were isolated from the different tissues using TRIzol reagent (Invitrogen) by a method previously described [20]. After treatment with DNase I, the first-strand cDNA was synthesized by reverse transcriptase SuperScript III (Invitrogen) with Random Hexamer Primer (Fermentas).…”
Section: Tissue Distribution Of Mjarf1 and Mjarfn Mrna Expressionmentioning
confidence: 99%
“…But then, the pathway of secretion or endocytic is easy to be employed to cause viral immune evasion and virus propagation, which Arfs was recruited to the specific target membrane to induce the disintegration of intracellular membranes and formation of specific vesicles by viral proteins mediating their interactors [18,19]. Moreover, Mjcent [20], as the Arf6 GAP, and MjArf4 [21], the member of Arf class II in Marsupenaeus japonicus, were up-regulated after WSSV infection, suggesting that shrimp Arfs may also participate in virus invasion and/or host defense events.…”
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