Cloning and sequence analysing of cytochrome P450 1A gene from white cloud mountain minnow Tanichthys albonubes
WANG Fang1,2, LI Kai-bin1, NIE Xiang-ping3, WU Shu-qin1
1. Pearl River Fisheries Research Institute, CAFS. Guangzhou 510380, China; 2. Shanghai Fisheries University, Shanghai 200090, China;3. Institute of Hydrobiology, Ji'nan University, Guangzhou 510632, China
The fish cytochrome P450 1A(CYP1A) gene is a useful biomarker in assessing contamination of the aquaticenvironment.In this paper,the CYP1A cDNA sequence of white cloud mountain minnow Tanichthys albonubes wereinvestigated.Total RNA was isolated from white cloud mountain minnow treated with β-NF,the cDNA of white cloudmountain minnow CYP1A gene were obtained using RT-PCR and RACE(rapid amplification of cDNA ends) PCRamplification from total RNA.The results of sequence analysis indicated that the white cloud mountain minnow CYP1A cDNAhas 2177bp,containing an open reading frame(ORF) of 1566bp encoding a 521 amino acids protein which yields a proteinmolecular weight is 58.5kDa.The obtained sequence possesses all characteristic motifs of teleost CYP1A genes including thestart codon,heme-binding cysteine(position 456-465),arginine codon(position 281) integral to enzymatic function and stopcodon(position 522).Alignment of the deduced amino acids of CYP1A cDNAs showed 87.7%、70.1%、76.2%、62.2%、54.5%、55.7% identities with Danio rerio,Fundulus heteroclitus,Oncorhynchus mykiss,Microgadus tomcod,Homo sapiens,Musmusculus,respectively.
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