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目的克隆血糖调节基因。方法按本室以往建立的方法制备大鼠血糖自动调节模型,以灌输生理盐水为对照;取其骨骼肌,采用差异显示技术(DDRT-PCR),获得差异表达基因片段,经狭缝杂交和Northern杂交分析,排除假阳性片段,确定真正的差异表达序列标签(EST),并以其为探针筛选大鼠骨骼肌cDNA文库。结果获得一个新的全长cDNA,命名为Fang-2,GenBank收录号为AF399874。经采用Blast软件(NCBI)分析显示,Fang-2是人类troponinT在大鼠中的同源物,其同源性为78%,表明该家族蛋白具有保守性。在高浓度的葡萄糖刺激大鼠后,和对照大鼠相比,Fang-2的表达下降。结论从大鼠骨骼肌中克隆到一个参与血糖调节的新基因,该基因可能通过其表达下调控制或部分控制某些目前未知的机制而达到调节血糖水平的作用。
Objective cloning blood glucose regulation gene. Methods The automatic glucose regulation model was established according to the method previously established in our laboratory. The rats were infused with normal saline as control. The skeletal muscle was harvested and the differentially expressed genes were obtained by differential display (DDRT-PCR) Hybridization analysis was performed to exclude false positives, and the real differentially expressed sequence tag (EST) was determined. The rat skeletal muscle cDNA library was screened by using it as a probe. The results obtained a new full-length cDNA, named Fang-2, GenBank accession number AF399874. Analysis using Blast software (NCBI) showed that Fang-2 is a homologue of human troponinT in rats with a homology of 78%, indicating that this family of proteins is conserved. After high concentration of glucose stimulated rats, Fang-2 expression decreased compared with control rats. Conclusion A novel gene involved in blood glucose regulation is cloned from rat skeletal muscle. The gene may regulate blood sugar level by regulating its expression or partially controlling some currently unknown mechanisms.