文献类型：期刊文献

中文题名：马氏珠母贝贝壳基质蛋白基因Pm-PNU7的克隆和功能研究

英文题名：Cloning and Functional Analysis of the Matrix Protein Gene Pm-PNU7 from the Pearl Oyster Pinctada martensii

作者：黄荣莲[1];范闪闪[1];杜晓东[1]

机构：[1]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,广东湛江524088

年份：2019

卷号：39

期号：1

起止页码：22

中文期刊名：广东海洋大学学报

外文期刊名：Journal of Guangdong Ocean University

收录：CSTPCD

基金：广东省科学技术厅(2017A030303076;2017A030307024);国基自然科学基金(31672626)

语种：中文

中文关键词：马氏珠母贝;贝壳基质蛋白;原位杂交;RNA干扰

外文关键词：Pinctada martensii;shell m atrix proteins;in situ hybridization;RNA interference

中文摘要：【目的】研究马氏珠母贝(Pinctada martensii)贝壳基质蛋白(Shell Matrix Proteins, SMPs)基因(Pm-PNU7)表达与功能。【方法】利用马氏珠母贝基因组和蛋白质组信息分析和RACE技术克隆获得壳基质蛋白新基因Pm-PNU7全长序列,通过原位杂交对其进行定位,RNA干扰检测其对贝壳形成的影响。【结果】Pm-PNU7的cDNA全长为797 bp,编码145个氨基酸,具有特殊的"KGG"重复序列和富含天冬氨酸(Asp)序列"DDDDDDHDD"。qRT-PCR分析表明,Pm-PNU7基因在外套膜边缘区和套膜区显著高表达(P<0.05);ISH检测显示,Pm-PNU7主要定位于边缘区外褶外侧和内侧上皮细胞、中褶内侧上皮细胞以及套膜区外侧上皮细胞。RNA干扰后,Pm-PNU7在外套膜边缘区和套膜区的表达量均显著下调,贝壳棱柱层和珍珠层微观结构均出现不同程度的紊乱。【结论】Pm-PNU7参与了贝壳棱柱层和珍珠层的形成。

外文摘要：【Objective】The expression and function of a novel shell matrix protein Pm-PNU7 in pearl oyster Pinctada martensii were studied according to the genome and proteome analyses.【Method】The full-length sequence of Pm-PNU7 gene of was cloned by RACE technology.Its transcripts was localized by in situ hybridization and the function of the gene in shell formation was studied by RNA interference.【Results】The full-length SMP Pm-PNU7 gene was 797 bp,encoding a novel protein with 145 amino acids with no functional annotation.It contained a special repeating sequence“KGG”and an aspartic acid-rich region“DDDDDDHDD”.qRT-PCR analysis results showed that Pm-PNU7 was significantly highly expressed in mantle edge(ME)and mantle pallial(MP)(P<0.05).ISH results showed that Pm-PNU7 mainly distributed in the inner and the outer epidermal cells of the outer fold of ME,the inner epidermal cells of the middle fold of ME,and the outer epidermal cells of MP.After gene knock down of Pm-PNU7 in ME and MP(P<0.05)by RNAi,,the microstructure of both prismatic layers and nacreous layers appeared distorted【Conclusion】Pm-PNU7 could affect the formation of the prismatic and nacreous layers of P.martensii.