条带杂交mRNA定量分析及其在血栓调节蛋白基因表达研究中的应用  

QUANTITATIVE ANALYSIS OF mRNA BY SLOT BLOT AND ITS APPLICATION TO THE STUDY OF GENE EXPRESSION OF THROMBOMODULIN

作　　者：周泉生[1] 储小红[1] 顾建明[1] 王晓冬[1] 阮长耿[1] 

机构地区：[1]江苏省血液研究所 苏州医学院血栓与止血研究室,苏州215007

出　　处：《中国实验血液学杂志》1994年第4期423-426,共4页Journal of Experimental Hematology

摘　　要：含血栓调节蛋白cDNA的质粒转染大肠杆菌,经扩增、酶切、凝胶电泳纯化,获得长度为3.7kb的血栓调节蛋白cDNA,后者用随机引物法标记^(32)P,制成血栓调节蛋白的基因探针。将该探针与血栓调节蛋白mRNA进行条带杂交后,分别用β-液闪仪和放射自显影及密度扫描仪作定量分析。用本法研究了血管内皮细胞中血栓调节蛋白基因表达及血小板血栓调节蛋白mRNA的含量。A TM cDN A (3,7kb) from pUC plasmid digested with EcoR I and recovered for agarose gel was labelled with 32P-dCTP as the probe for quantitative detection of TM mRNA in vascular endothelial cells and platelets by means of a Slot-blotting. The results showed that 106 endothelial cells contained 2.7 ± 0.3ng TM mRNA; the amount of TM mRNA in 5 × 108platelets is was 2.68± 0.25ng. Futhermore, these studies also indicated , that, new-breviscapine, dybutyl-cAMP and defibrotide could increase the mRNA levels of TM in endothelial cells; but, tetramethylpyrazine had no effect on the expression of TM mRNA; in contrast, it decreased the levels of TM mRNA.

关 键 词：血栓调节蛋白mRNA 基因表达 血管内皮细胞 血小板 

分 类 号：R346[医药卫生—基础医学]