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Cloning and expression of group IB phospholipase A 2 isoforms in the red sea bream, Pagrus major
Author(s) -
Iijima N.,
Fujikawa Y.,
Tateishi Y.,
Takashima Y.,
Uchiyama S.,
Esaka M.
Publication year - 2001
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-001-0749-x
Subject(s) - pagrus major , gene isoform , lipidology , clinical chemistry , cloning (programming) , fishery , pagrus , biology , zoology , chemistry , gene , biochemistry , fish <actinopterygii> , computer science , programming language
Two cDNA encoding red sea bream DE‐1 and DE‐2 phospholipases A 2 (PLA 2 ) were cloned from the hepatopancreas of red sea bream, Pagrus (Chrysophrys) major . The cDNA of DE‐1 PLA 2 encoded a mature protein of 125 amino acid residues with an apparent signal peptide of 20 residues and propeptide of 5 residues, and that of DE‐2 PLA 2 , a mature protein of 126 amino acid residues with an apparent signal peptide of 17 residues and propeptide of 6 residues. Comparison of the predicted amino acid sequences for mature DE‐1 and DE‐2 PLA 2 showed that both proteins contain 14 cysteines including Cys 11 and 77 and a pancreatic loop, which are commonly conserved in group IB PLA 2 ; however, the identity in amino acid sequence between DE‐1 and DE‐2 PLA 2 was low (47%). A previous report concerning the cDNA cloning of red sea bream gill G‐3 PLA 2 and the present results represent the first cloning and sequencing of three distinct isoforms of group IB PLA 2 in a single fish species, red sea bream. Reverse transcription‐polymerase chain reaction analysis showed that DE‐1 PLA 2 mRNA was expressed in the hepatopancreas, pyloric ceca, intestine, spleen, gonad, stomach, and kidney, whereas gill G‐3 PLA 2 mRNA was expressed only in the gills and gonad. The expression of DE‐2 PLA 2 mRNA was detected in all of the tissues analyzed. These results indicate that three distinct isoforms of group IB PLA 2 , DE‐1 and DE‐2 PLA 2 in hepatopanceas and gill G‐3 PLA 2 , are expressed in a tissue‐specific manner in red sea bream.