cDNA cloning and expression of Acyl-CoA binding protein (ACBP) from orange-spotted grouper, <i>Epinephelus coioides</i><i><span style="font-family:"Book Antiqua";mso-bidi-font-family:Arial" lang="EN-GB"></span></i>

Abstract

cDNA of acyl-CoA binding protein (ACBP) was constructed, cloned and expressed in hypothalamus and pituitary of orange-spotted grouper, <i>Epinephelus coioides.</i> <span style="mso-bidi-font-style: italic">The ACBP gene contains 630 nucleotides including the open reading frame (ORF) of length 128 bp. The open reading frame was started with ATG codon at the position 462 and terminated with a stop codon TAA at the position 600. The ORF sequence consisted of 42 amino acids and the calculated mol wt of the protein was approx 4.62 kDa (GenBank: FJ939282.1). The highest identity was observed with the puffer fish, <i>Tetraodon nigroviridis</i> and <i>Anoplopoma fimbria</i> with homology of 83 and 88.1%, respectively. RT-PCR results revealed that ACBP mRNA was abundantly expressed in pituitary, hypothalamus and medulla oblongata. Further, ACBP was cloned in pGEX-KG expression vector and transformed into BL21(DE3). The protein was purified by affinity chromatography and partially characterized. <span style="mso-bidi-font-weight:bold">The abundant expression of ACBP in pituitary and hypothalamus, in the <span style="mso-bidi-font-weight: bold">present study, indicated that it might be involved in reproduction.

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