摘要
β—淀粉样蛋白前体(APP)与tau蛋白是两种与Alzheimer病的病理改变相关的蛋白质,近来也发现它们或其修饰产物在包含体肌炎及慢性氯喹中毒的肌细胞中沉积。为更好地研究这两种相关蛋白质在肌细胞中的表达,我们建立了用逆转录-多聚酶链反应同时定量这两种基因转录产物的方法。本法选用甘油醛-3-磷酸脱氢酶(G_3PD)作为内标,用激光图像分析测定扩增产物。结果显示,PCR在很宽的范围内呈指数扩增,两种靶mRNA与内标mRNA扩增效率相当,因而可以G_3PD来校正APP与tau的测定结果。初步实验显示氯喹中毒首先促进肌肉细胞中APP与tau蛋白的表达,在第4周达到高峰,然后表达逐渐减少,在第10周转为明显抑制作用。
We describe here a method for simultaneous relative quantitation of APP and tau expressions in rat muscle using polymerase chain reaction combined with reverse transcription. The target mRNAs were coamplified with an internal control, glyceraldehyde-3-phosphate dehydrogenase (G,PD). The amplified products were measured with a laser image analyzer,and the values were plotted against the cycle number. In the exponential phase of amplification, the linearity and re-producibility of RT-PCR demonstrate identical efficiencies of cDNA synthesis and PCR amplification for APP,tau and G3PD. These results suggest that in the exponential phase of amplification, a relative quantitation of target mRNAs could he accomplished by normalization with internal control. Preliminary experiments indicate that the expressions of both APP and tau in rat muscle were first increased and then decreased by chronic intoxication with chloroquine.
出处
《实验生物学报》
CSCD
1998年第1期1-6,共6页
Acta Biologiae Experimentalis Sinica
关键词
早老性痴呆
淀粉样蛋白前体
TAU蛋白
逆转录
Alzheimer's disease. β-Amyloid protein precursor. Tau protein.Chloroquine. mRNA quantita-tion.
