Zhou Yuchan, Kaye Peter L, Pantaleon Marie
Department of Physiology and Pharmacology, School Of Biomedical Sciences, The University of Queensland, Brisbane, Qld 4072, Australia.
Gene Expr Patterns. 2004 Oct;4(6):621-31. doi: 10.1016/j.modgep.2004.04.010.
In this study we report the cloning and characterisation of the mouse Glut12 gene and examine for the first time its expression pattern in the earliest stages of development. Mouse Glut12 (mGlut12) was cloned from preimplantation embryos by 5'RACE RT-PCR using primers designed from an EST clone corresponding to a human GLUT12 antigenic sequence after positive immunoreactivity was observed in mouse two-cell embryos by western immunoblotting. The mGlut12 gene contains an open reading frame of 1869 base pairs, potentially encoding a polypeptide of 622 amino acids. The predicted mGLUT12 protein bears all the hallmarks of the SLC2A family of hexose transporters and shares an 83% sequence homology to human GLUT12. Consistent with its human homolog mGlut12 mRNA is found highly expressed in skeletal and cardiac muscle and fat. Additionally, it was also found in the uterus and during early embryogenesis. During early development in the mouse, Glut12 expression is clearly apparent in ovulated oocytes and two-cell embryos but declines in day 3 morulae. With the exception of some Glut12 expression apparent in blastocysts, Glut12 mRNA remains at low to undetectable levels until E11.
在本研究中,我们报告了小鼠Glut12基因的克隆与特性分析,并首次检测了其在发育早期阶段的表达模式。通过5'RACE RT-PCR从植入前胚胎中克隆小鼠Glut12(mGlut12),所用引物是根据一个与人类GLUT12抗原序列对应的EST克隆设计的,此前通过western免疫印迹在小鼠二细胞胚胎中观察到阳性免疫反应。mGlut12基因包含一个1869个碱基对的开放阅读框,可能编码一个622个氨基酸的多肽。预测的mGLUT12蛋白具有己糖转运蛋白SLC2A家族的所有特征,与人类GLUT12的序列同源性为83%。与其人类同源物一致,mGlut12 mRNA在骨骼肌、心肌和脂肪中高度表达。此外,在子宫和早期胚胎发育过程中也发现了它。在小鼠早期发育过程中,Glut12在排卵的卵母细胞和二细胞胚胎中表达明显,但在第3天的桑椹胚中表达下降。除了在囊胚中明显有一些Glut12表达外,Glut12 mRNA在E11之前一直处于低水平至无法检测的水平。