Expression and Functional Characterization of the RIG-I-Like Receptors MDA5 and LGP2 in Rainbow Trout (Oncorhynchus mykiss)

Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella

“…#FP512). In addition, the gcPGRP6 ORF was amplified using primers GFPPGRP6/pt1PGRP6R (Table 1) by PCR and inserted into the Sac I and Bam HI sites of ptGFP1 vector which contained two identical CMV promoters and SV40 3'UTRs to direct the expression of target gene and GFP gene independently in transfected cells (Chang et al, 2011).…”

Fish MAVS is involved in RLR pathway-mediated IFN response

“…Characterization of these genes individually suggests that they all are involved in fish IFN response. For example, overexpression of salmon RIG-I or rainbow trout MDA5 results in upregulation of IFN and ISGs and also establish host antiviral states [21,22]; ectopic expression of crucian carp IRF3 or MITA triggers the expression of both IFN and ISGs [23,24]. Fish MAVS genes are first identified from Atlantic salmon (Salmo salar) and zebrafish (Danio rerio), both of which exhibit an ability to induce transcriptional expression of IFN and confer full protection against both RNA and DNA virus on cultured fish cells [21,25].…”

Expression and functional characterization of PGRP6 splice variants in grass carp Ctenopharyngodon idella

“…For overexpression studies of gcPGRP6 variants, the modified expression vector ptGFP1 was used (Chang et al, 2011), which contained two sets of CMV promoters and SV40 3' UTR, to drive expression of the target gene product and GFP as separate proteins rather than as a fusion protein. The cDNAs of gcPGRP6 variants were amplified with primer pairs GFPPGRP6F/pt1PGRP6R and GFPPGRP6F/pt1PGRP6Ra, and inserted into the Sac I and Bam HI sites of ptGFP1 vector with a stop codon to generate ptGFP1-gcPGRP6, ptGFP1-gcPGRP6a, ptGFP1-gcPGRP6b, ptGFP1-gcPGRP6c and ptGFP1-gcPGRP6d expression plasmids (Supplementary Table S1).…”