马氏珠母贝载脂蛋白-2基因的克隆与表达分析

摘要

The full length of Pm-ApoL2 was cloned from Pinctada fucata martensii by RACE technology. The traditional extraction method was applied to separately extract total carotenoid content of yellow and white colored adductor samples. The expression of Pm-ApoL2 in tissue-specific and yellow and white colored adductor samples was detected by real-time PCR. The results showed that the total length of Pm-ApoL2 gene was 1328 bp,including 705 bp of the open reading frame (ORF) that encoded 234 amino acids, a 5′UTR of 342 bp and a 3′ UTR of 281 bp. Fluorescence quantitative PCR analysis showed that the hepatopancreas had the highest expression of Pm-ApoL2 gene, followed by mantle, gill and adductor muscle. The gene expression was significantly higher in the hepatopancreas than in the other three tissues (P < 0.05). Yellow colored adductor samples had significantly higher total carotenoid content than white colored adductor samples (P < 0.05). Yellow colored adductor samples had significantly higher gene expression than white colored adductor samples (P < 0.05). The results suggested that Pm-ApoL2 was a candidate gene involved in carotenoid metabolism in pearl oyster P. fucata martensii.%以RACE技术克隆获得马氏珠母贝载脂蛋白-2(Pm-ApoL2)基因cDNA全长序列,以传统萃取法提取黄色与白色闭壳肌个体的类胡萝卜素,以荧光定量技术检测Pm-ApoL2基因在各个组织中的表达量及黄与白闭壳肌个体的Pm-ApoL2表达量.结果表明,Pm-ApoL2基因cDNA序列全长1328 bp,开放阅读框(ORF)长705 bp,编码234个氨基酸,5′ 非翻译区(5′UTR)长342 bp,3′UTR长281 bp.Pm-ApoL2在肝胰腺中表达量最高,其后依次是外套膜、鳃、闭壳肌,其中肝胰脏表达量显著大于其它3个组织(P<0.05).黄、白色闭壳肌个体之间的类胡萝卜素含量和Pm-ApoL2的表达量都存在显著性差异(P<0.05),且二者的类胡萝卜素含量与Pm-ApoL2表达量呈显著正相关.Pm-ApoL2基因为马氏珠母贝类胡萝卜素代谢候选基因.