Fructosel, 6-bisphosphatase ( EC 3. 1. 3. 11) can catalyse the hydrolysis of fructose-l, 6-bisphosphate into fructose-6-phosphate and inorganic phosphate. It is one of the key enzymes in the gluconeogenic pathway. The full-length 1 357 bp FBP cDNA was isolateded from the liver of sea perch Lateolabrax japonicus with SMART RACE technology. It consists of a 42 bp5' UTR,a 301 bp 3' UTR and an open reading frame (ORF) of 1 014 bp,encoding a polypeptide of 337 amino acids with a predicted molecular weight of 36. 7 kD and electric point of 6. 90. Sequencing analysis of amino acid shows that the Lateolabrax japonicus FBP has high amino acid homology with the liver-type FBP gene from other species, and the homology is 94. 3% with Anopiopoma fimbria, 90. 8% with Osmerus mordax,89. 3% with Danio rerio, 88. 1% with Carassius gibelio and 84. 1% with Salmo salar. Phylogenetic analysis shows that Lateolabrax japonicus FBP clustered with liver-type FBP of fish, then with liver-type FBP of mammals, and still then clustered with muscle-type FBP of mammals to form a family. Semi-quantitative RT-PCR was used to determine the expression profiles of FBP from liver, muscle, heart, eye,intestine, kidney, fat, spleen, gill and brain. The results show that FBP was expressed only in liver, intestine and kidney. This conclusion is in consistent with the fact that liver, kidney and intestine are the major gluconeogenic tissues of teleost. These results indicate that the FBP obtained is of liver-type.
Key words
Fructose-1,6-bisphosphatase /
Lateolabrax japonicus /
cloning /
expression
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