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。
【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.
作者
黄荣莲
范闪闪
杜晓东
HUANG Rong-lian;FAN Shan-shan;DU Xiao-dong(Fisheries College,Guangdong Ocean University//Pearl Breeding and Processing Engineering Technology Research Center of Guangdong Province,Zhanjiang 524088,China)
出处
《广东海洋大学学报》
CAS
2019年第1期22-29,共8页
Journal of Guangdong Ocean University
基金
广东省科学技术厅(2017A030303076
2017A030307024)
国基自然科学基金(31672626)
关键词
马氏珠母贝
贝壳基质蛋白
原位杂交
RNA干扰
Pinctada martensii
shell m atrix proteins
in situ hybridization
RNA interference
