摘要
Apolipoprotein L3(APOL3)是新发现的一种高密度脂蛋白,属于载脂蛋白L基因家族的成员之一,具有运载脂类物质进行生化代谢的功能。本研究利用c DNA末端快速扩增技术获得了马氏珠母贝APOL3基因(Pm-APOL3)c DNA全长序列,分析其序列及功能特征,检测Pm-APOL3基因在不同组织的表达,并分析了金黄壳色群体与养殖群体类胡萝卜素含量与Pm-APOL3基因表达量的相关性。结果表示,Pm-APOL3基因c DNA全长874 bp,开放阅读框(ORF)为357 bp,编码118个氨基酸,包含475 bp的5'UTR,42 bp的3'UTR,相对分子量为12.795 5 ku,等电点为4.71。该蛋白质的不稳定指数为34.09,属于稳定蛋白质,疏水性分析表明其亲水性平均系数为0.74,属疏水蛋白,信号肽预测结果表示其不含信号肽,为非分泌蛋白。荧光定量PCR分析表明Pm-APOL3基因在鳃、外套膜、闭壳肌与肝胰脏均有表达,其中肝胰脏表达量显著大于其它三个组织(p<0.05)。金黄壳色群体类胡萝卜素含量与Pm-APOL3基因表达量均显著大于养殖群体(p<0.05)。本研究结果表明Pm-APOL3是参与马氏珠母贝类胡萝卜素代谢重要基因。
Apolipoprotein L3, a kind of high density lipoprotein, is a newly discovered member of the apolipoprotein L gene family, which is involved in lipid biochemical metabolism. By using rapid amplification of c DNA ends technology, we cloned the full-length sequence of APOL3 gene of Pinctada fucata martensii(Pm-APOL3), analyzed functional characteristics, detected the relative expression in the tissues, and estimated the relationship between the gene relative expression and total carotenoid. The results showed that the Pm-APOL3 gene full-length was 874 bp, which contained an open reading frame of 357 bp that encoded 118 amino acids, with5' UTR of 475 bp and 3' UTR 42 bp. The relative molecular weight was 12.795 5 ku and isoelectric point was 4.71.The protein instability index was 34.09, which belongs to the stable protein. The hydrophilic average coefficient was 0.74, which belongs hydrophobic protein, and the signal peptide prediction result showed that it is a secreted protein, because it contained no signal peptide. Fluorescence quantitative PCR analysis showed that Pm-APOL3 gene could be detected in gill, mantle, adductor, and hepatopancreas. The gene expression was significantly higher in the hepatopancreas than the other three tissues(p<0.05). Total carotenoid and gene expression were significantly larger in the yellow colored stock than in the cultured stock(p<0.05). The present results indicated that Pm-APOL3 was involved in transport of carotenoid metabolism in pearl oyster Pinctada fucata martensii.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2016年第7期1707-1714,共8页
Genomics and Applied Biology
基金
广东省科技厅项目"近江牡蛎三倍体优质苗种及马氏珠母贝新品种培育技术研究与开发"(2013B020201002)
广东省海洋与渔业局科技专项"马氏珠母贝优质抗逆选系苗种规模化繁育与示范养殖"(Z2014009)
2015年广东海洋大学"海之帆-起航计划"大学生科技创新项目"马氏珠母贝类胡萝卜素与相关基因表达量在关联分析"
2015年广东海洋大学水产学院创新项目"马氏珠母贝类胡萝卜富集机理分析"共同资助
关键词
马氏珠母贝
载脂蛋白L3
基因克隆
类胡萝卜素
Pinctada fucata martensii
Apolipoprotein L3
Gene cloning
Total carotenoid
