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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 that plays an important role in mediating the regulation of calcium-phosphorus metabolism in the body, participation in bone formation and mineralization, and bone metabolism. In this experiment, we obtained the coding region of vitamin D receptor (Pm-VDR) from Pachytene marxo and detected the expression pattern of Pm-VDR in different tissues. We also analyzed the expression of Pm-VDR and P - type voltage-dependent calcium channel (PCaβ) gene expression to explore Pm-VDR in the bio-mineralization of Pinctada martensii role. The results showed that the open reading frame (ORF) of Pm-VDR was 1407bp and encoded 468 amino acids. The relative molecular mass was 53871.17kD and the isoelectric point was 6.19. The liposolubility index was 68.14 and the average hydrophilicity was -0.70. Aqueous protein. Pm-VDR has a DNA binding domain composed of two zinc finger structures and a ligand binding domain of a hormone receptor. Multiple sequence alignment showed that Pm-VDR was highly conserved among species and had the highest homology with long-oyster. qRT-PCR analysis showed that the expression of Pm-VDR in hepatopancreas and gill of pearl oyster was significantly higher than that of other tissues, of which hepatopancreas was the highest. The expressions of Pm-VDR and PCaβ in the gill were significantly up-regulated (P <0.05) after 0.2%, 2%, 20% of VD3 stimulation for 12h, and the minimum concentrations were up to 2% and 0.2%, respectively. Taken together, Pm-VDR may regulate the absorption of Ca2 + by regulating the expression of PCaβ gene by VD3.