企鹅珍珠贝GLUT1基因全长cDNA克隆及其对葡萄糖的表达响应

Cloning of GLUT1 gene from winged pearl oyster Pteria penguin and its expression in response to glucose challenge

  • 摘要: 葡萄糖转运蛋白(GLUTs)是能量代谢的关键因子,为了探索GLUT1(glucose transporter type1)在企鹅珍珠贝(Pteria penguin)中的功能,开展了企鹅珍珠贝GLUT1基因的克隆与功能研究。利用RACE技术克隆出企鹅珍珠贝GLUT1基因,并命名为PpGLUT1。该基因cDNA全长为2 068 bp, 编码522个氨基酸,有2个跨膜螺旋区,N端和C端都在胞质侧,分子量为57.226 4 kD,理论等电点为5.17。不同组织荧光定量结果表明,在肠和外套膜中表达量最高,闭壳肌中的表达量显著低于其他组织。0.5 g·mL-1的高质量浓度葡萄糖注射实验表明,在注射后30 min内,肠PpGLUT1表达量升高,在30 min之后表达量逐渐降低,注射后第2小时表达量最低。不同浓度葡萄糖注射实验表明,当葡萄糖质量浓度为0.1~0.4 g·mL-1时,PpGLUT1表达量逐渐上升;质量浓度为0.4 g·mL-1时,PpGLUT1表达量最高;大于0.4 g·mL-1之后,表达量逐渐降低。上述结果表明,企鹅珍珠贝PpGLUT1主要在肠和外套膜表达,并对葡萄糖注射有表达响应,可能在上述组织的葡萄糖转运过程中发挥作用。

     

    Abstract: Facilitated glucose transporter type 1 (GLUT1) takes part in transporting glucose in most animal tissues responsible for energy metabolism. To study function of GLUT1 in Pteria penguin, the cDNA sequence of GLUT1 was obtained from P.penguin transcriptome using RACE, which was named as PpGLUT1. The full length cDNA was 2 068 bp, The gene encoded 522 amino acids, 12 transmembrane helical motifs and multiple phosphorylation sites, with N-terminal and C-terminal on the cytoplasmic side.The molecular weight was 57.226 4 kD and the isoelectric point was 5.17. Fluorescent quantitative PCR results show that PpGLUT1 expressed in all tissues and was significantly low in muscle. The high concentration of 0.5 g·mL-1 glucose injection result shows that the expression of PpGLUT1 increased within 30 min after injected, and then decreased; at the injection time of 2 h, the expression of PpGLUT1 was lowest; the expression gradually increased after 2 h. Different concentrations of glucose injection test indicate that the expression levels of PpGLUT1 increased with increase of glucose concentrations ranging from 0.1 g·mL-1 to 0.4 g·mL-1 and then decreased. The mRNA expression responses to glucose suggests that PpGLUT1 plays an important role in glucose transportation in multiple tissues of P.penguin.

     

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