马氏珠母贝STARDL3基因的克隆与表达性分析.docVIP

马氏珠母贝STA RDL3基因的克隆与表达 性分析 雷超郑哲王庆恒黄荣莲邓岳文杜晓东 广东海洋大学水产学院广东省珍珠养殖与加工工程 技术研究中心 采用RACE技术克隆获得马氏珠母贝STARDL3基因c DNA全长序列(Pm-STARDL3); 利用荧光定量技术检测Pm-STARDL3基因在各个组织中的表达量。结果表明， Pm-STARDL3基因c DNA序列全长3 655 bp,开放阅读框(ORF)长1 491 bp,编 码 496 个氨基酸，5’非翻译区(5’UTR)长 110 bp, 3’UTR 长 2 054 bp。PmSTARDL3 氨基酸序列同源比对分析显示与华贵类捞孔扇贝(Mimachlamys nobilis) STARDL3的序列的相似度最高。SMART软件分析得出，Pm_STARDL3有STARD类蛋 白特有的结构域。荧光定量PCR检测结果表明Pm-STARDL3在肝胰腺中表达量最 高，其后依次是外套膜、鳃与闭壳肌，各组织的表达量差异具统计学意义 (p0. 05)。 关键词： 马氏珠母贝;STARDL3;基因兗隆;基因表达; 邓岳文，dengyw@gdou. 基金：国家自然基金“马氏珠母贝生长性状QTLs精细定位与相关基因功能分析 ” Clone and Expression Analysis of STARDL3 Gene from Pinctada fucata martensii Lei Chao Zheng Zhe Wang Qingheng Huang Ronglian Deng Yuewen Du Xiaodong Fisheries College, Guangdong Ocean University; Abstract： Tn this study, the full length of Pm-STARDL3 was obtained by using RACE technology. The expression of tissue-specific was detected by Real-time 1)CR. The results showed that the total length of l)m-STARDL3 gene was 3 655 bp, including 1 491 bp of the open reading frame (ORF) that encoded 496 amino acids, a 5’UTR of 110 bp and a 3’ UTR of 2 054 bp. Multiple sequence alignment indicated that Pm-STARDL3 was high identity with Mimachlamys nobilis. Protein sequence analysis showed that Pm-STARDL3 has a typical START structural domain.Fluorescence quantitative PCR analysis showed that the hepatopancreas had the highest expression of Pm_STARDL3 gene, followed by mantle, gill and adductor muscle, and the expression difference of the organization had statistical significance. Keyword： Pinctada fucata martensii; STARTDL3; Gene cloning; Gene expression; st AR 相关脂质转运蛋白-3 类似蛋白(st AR-related lipid transfer protein 3-like protein, STARDL3)属于类固醇急性调节蛋白(steroidogenacute regulatory St AR)家族。它与 STARD3 (st AR-related lipid transferprotein 3)有很高的同源性，都有START结构域，此结构域是STARD类蛋Q的特有结构 域(邢晋祎等，2015),能结合多个疏水配体并经常出现在多结构域蛋白中，起 到调节其它功能结构域的作用(Roderick et al., 2002),因而参与众多的生 理过程。目前己知STARD3参与肭固醇、类固醇和脂类的物质的合成以及代谢过 程(裘越，2013)。例如，人类视网膜STARD3是类胡萝卜素结合蛋闩，其作用 是特异性结合叶黄素(H