cDNA Sequence Cloning and Tissue Expression of Uncoupling Protein 2 of Silver Carp (Hypophthalmichthys molitrix)
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Abstract
PCR using two degenerated primers designed on the basis of the conserved region of mammals and fish uncoupling protein 2 (UCP2) gene yielded a cDNA fragment of 620 bp in the liver of silver carp. This cDNA fragment was completed by 5′ and 3′ RACE. The full-length silver carp UCP2 cDNA clone was 1 452 bp in length, containing an open reading frame of 933 bp (encoding 310 amino acids), flanked by 337 bp 5′UTR and 182 bp 3′UTR. The deduced amino acid sequence from this UCP2 cDNA fragment contains three signature motifs of mitochondrial transporter protein, and UCP-specific sequences. Tissue-expression analysis revealed that the silver carp UCP2 was highly expressed in intestine, liver, muscle and adipose tissue, but lower in brain tissue. This tissue expression pattern is consistent with the microcystins distribution among these tissues, suggesting that UCP2 expression level has a close relationship with the overproduced ROS which is induced by microcystins.
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