大黄鱼CYP1A基因cDNA的克隆与表达分析

Cloning and Expression Analysis of CYP1A Gene from Larimichthys crocea

由于外源化合物能诱导鱼类CYP1A(P4501A)的表达,因而它广泛被用作评价水环境污染生物标记物。利用RT-PCR结合RACE技术从大黄鱼(Larimichthys crocea)肝脏克隆了CYP1A基因全长cDNA序列。经分析,该cDNA的5'末端有175 bp的非翻译区,开放阅读框为1 566 bp,编码521个氨基酸和一个终止密码子,3'末端有857 bp的非翻译区,3'非翻译区有一个多聚腺苷酸信号及两个与mRNA的快速降解有关的AUUUA序列。推测大黄鱼CYP1A的氨基酸序列和欧洲鲈鱼的相似度最高达89.6%。用RT-PCR检测大黄鱼CYP1A的表达特征发现,在所检测的9个组织中均有表达,以肝脏、消化道、脾脏和肾脏的表达量较高。

The induction of fish CYP1A by various xenobiotics has led to its use as a biomarker in assessing contamination of the aquatic environment.We have cloned a full-length cDNA of CYP1A gene using reverse transcription polymerase chain reaction（RT-PCR）and rapid amplification of cDNA ends（RACE）from the liver of large yellow croaker（Larimichthys crocea）.The obtained cDNA contained a 5′ untranslated region of 175 bp,an open reading frame of 1 566 bp coding for 521 amino acids and a stop codon,followed by a 3′ untranslated region of 857 bp.The 3′ UTR contained poly-adenylation signal and two AUUUA sequences that are likely to be a target for rapid degradation of its mRNA.The deduced amino acid sequence of Larimichthys crocea CYP1A shows 89.6% identity with that of Dicentrarchus labrax.CYP1A gene was expressed in all 9 tissues tested by RT-PCR and the highest expression was found in liver,intestine,kidney and spleen.