CDNA for an immune response gene encoding low molecular weight polypeptide from flounder, Paralichthys olivaceus

The cDNA for an immune response gene encoding the low molecular weight polypeptide (LMP7) was cloned and sequenced from a flounder (Paralichthys olivaceus) leukocyte cDNA library. The cDNA clone was 1,160 bp, and composed of an open reading frame of 822 bp that corresponded to a protein of 273 amino acid residues with a calculated mass of 30.5 kDa. The ScanProsite search indicated that the deduced amino acid sequence from the flounder LMP7 contains a proteasome beta-type subunit signature, which is well conserved during evolution. The sequence shares a high degree of identity with other LMP7 sequences varying from a 66% identity with zebra fish (Danio renio) to a 57% identity with the African clawed frog (Xenopus laevis), which was confirmed from a phylogenetic tree. A reverse transcription-polymerase chain reaction (RT-PCR) was used to determine tissue specificity, and the expression of LMP7 was detected from the liver, kidney, leukocyte, and spleen of the flounder.