摘要
维生素D受体(VDR)是一种核激素受体,在介导维生素D调节机体钙-磷代谢、参与骨形成和矿化以及骨代谢等方面发挥重要作用。本实验克隆获得马氏珠母贝维生素D受体(Pm-VDR)编码区,检测其在不同组织中的表达模式,并分析维生素D3(VD3)刺激后Pm-VDR和马氏珠母贝L-型电压依赖性钙通道(PCaβ)基因的表达,探究Pm-VDR在马氏珠母贝生物矿化过程中的作用。结果显示:Pm-VDR开放式阅读框(ORF)为1407bp,编码468个氨基酸;相对分子量为53871.17kD,等电点为6.19;脂溶性系数为68.14,总平均亲水性为-0.70,属于亲水性蛋白。Pm-VDR具有由两个锌指结构组成的DNA结合域和一个激素受体的配体结合域。多序列比对结果显示Pm-VDR在物种之间具有较高的保守性,与长牡蛎同源性最高。qRT-PCR分析发现Pm-VDR在马氏珠母贝肝胰腺和鳃中表达量显著高于其他组织,其中肝胰腺的表达量最高。在0.2%、2%、20%浓度的VD3刺激12h后,Pm-VDR和PCaβ基因在鳃中表达量均显著上调(p<0.05),上调的最低浓度分别为2%和0.2%。综上所述,Pm-VDR可能通过介导VD3调控PCaβ基因的表达参与马氏珠母贝鳃组织对Ca2+的吸收过程,从而调控贝体的生物矿化。
Vitamin D receptor （VDR） is a nuclear hormone receptor, which plays an important role in regulating the body＇s calcium-phosphorus metabolism, participating in bone formation and mineralization and bone metabolism. This study cloned the coding region of Pm-VDR and detected its expression pattern in different tissues. Also, we analyzed the expression of Pm-VDR and PCaβ after the stimulation of VD3 and explored the function of Pm-VDR in the biomineralization process of Pinctada martensi. The results showed that the open reading frame （ORF） of Pm-VDR cDNA was 1 407 bp, encoding 468 amino acids. The relative molecular weight was 53 871.17 kD and the isoelectric point was 6.19. The aliphatic index was 68.14 and the grand average of hydropathicity was -0.70, which meant it belonged to hydrophilic proteins. Pm-VDR had a DNA-binding domain with two zinc fingers and a ligand binding domain of hormone receptor. Multiple sequence alignment represented that Pm-VDR was highly conservative among species, and it was most homologous with Crassostrea gigas. In addition, qRT-PCR analysis indicated that the expression of Pm-VDR in the gill and hepatopancreas was significantly higher than that in other organizations, with the highest expression in the hepatopancreas. After the VD3 stimulation with 0.2%, 2% and 20% concentrations for 12 hours, the expression of Pm-VDR and PCaβ in gill was significantly up-regulated （p〈0.05）, and the minimum up concentration was 2% and 0.2%, respectively. In summary, VDR could regulate the expression of PCaβ by mediating VD3 and participate in the process of calcium absorption in P. f martensiii gill tissue, thereby regulating the biomineralization of shell.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2017年第7期2840-2848,共9页
Genomics and Applied Biology
基金
广东省普通高校重大科研项目培育计划"珍珠贝良种培育与新型育种模式"(2014GKXM049)
广东省海洋渔业与产业发展专项"马氏珠母贝优质抗逆选系苗种规模化繁育与示范养殖"(Z2014009)和"马氏珠母贝海选1号苗种规模化繁育与示范养殖"(B201601-Z10)共同资助
