Journal of Tropical Oceanography >
CYP4 gene cloning and expression level analysis of Perna viridis
Received date: 2008-05-26
Revised date: 2008-09-07
Online published: 2010-07-29
Supported by
科技部科研院所社会公益研究专项(2005DIB3J020); 中央级公益性科研院所基本科研业务费专项资金(中国水产科学研究院南
海水产研究所)资助项目(2007ZD08)。
In order to identify and obtain the cDNA sequence of CYP4 gene in Perna viridis, degenerate PCR and RACE (Rapid Amplification of cDNA Ends) were used, and partial CYP4 gene cDNA sequence total of 1197 bp was acquired (GeneBank/NCBI EU429566) from Perna viridis digestive gland. The CYP4 gene expression in digestive gland, foot and gonad of wild Perna viridis and the CYP4 gene expression level in gonad from Aroclor1254 treated Perna viridis were evaluated using real-time quantitative-PCR (RQ-PCR) and Sybr Green I chemistry. The RQ-PCR primers were designed based on the Partial cDNA sequences of Perna viridis CYP4, and the beta-actin target was used as internal reference and included in each RQ-PCR experiment for the normalization of expression data. The results showed that basic CYP4 expression existed in all of the three tissues (digestive gland, foot and gonad) from wild Perna viridis, and the expression of Perna viridis CYP4 had tissue differences and sex differences. The Perna viridis gonad CYP4 gene expression can be induced by Aroclor1254 exposure, and this induction is both exposure-time dependent and exposure-dose dependent. This study has provided useful clues to the inquisition of using CYP4 gene family as biomarker of toxicology effect.
Key words： CYP4 gene; Perna viridis; Quantitative-PCR
ZHOU Chi,LI Chun-hou,ZHANG Wei-min,JIA Xiao-ping . CYP4 gene cloning and expression level analysis of Perna viridis[J]. Journal of Tropical Oceanography, 2010 , 29(4) : 82 -88 . DOI: 10.11978/j.issn.1009-5470.2010.04.082
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