A novel immune-related gene was expressed in Japanese flounder （Paralichthys olivaceus） injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5＇UTR, a 336 bp open reading frame （ORF） encoding 111...A novel immune-related gene was expressed in Japanese flounder （Paralichthys olivaceus） injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5＇UTR, a 336 bp open reading frame （ORF） encoding 111 amino acids and a 556bp 3＇UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S （NMS）, but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa sig- nal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarurn. These results indicated that the PolR2 might play some important role in Japanese flounder immune response system. This gene was named PolR2 （P.olivaceus immune-related gene 2, GenBank accession number： EU224372）. The mature PoIR2 peptide was expressed in BL21 （DE3） pLysS using pET-32a（＋） vector and a great part of the recombinant mature peptide existed as soluble type.展开更多
基金supported by grants from the National High Technology Research and Development Program of China (Nos.2006AA10A404 and 2006AA10A414)
文摘A novel immune-related gene was expressed in Japanese flounder （Paralichthys olivaceus） injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5＇UTR, a 336 bp open reading frame （ORF） encoding 111 amino acids and a 556bp 3＇UTR. Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S （NMS）, but further alignment indicated they did not have NMS C-terminal conservational signature domain. So it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa sig- nal peptide and was a secretory pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after infected with V. anguillarurn. These results indicated that the PolR2 might play some important role in Japanese flounder immune response system. This gene was named PolR2 （P.olivaceus immune-related gene 2, GenBank accession number： EU224372）. The mature PoIR2 peptide was expressed in BL21 （DE3） pLysS using pET-32a（＋） vector and a great part of the recombinant mature peptide existed as soluble type.
