摘要
目的 :半定量检测体外培养的少量兔血管内皮细胞中特异mRNA的表达。方法 :用PharmaciaBiotech公司的快速微量mRNA提纯试剂盒从兔内皮细胞中直接提取出mRNA ,并以此为模板 ,用半定量RT -PCR技术检测培养的兔血管内皮细胞中内皮素 (ET - 1)mRNA的相对含量。结果 ;从 10 5个兔血管内皮细胞中提取出未降解的mRNA ,A2 6 0 /A2 80 >1.8;用半定量RT -PCR技术 ,测定出ET - 1mRNA的表达。结论 :本实验建立了一种从少量细胞中快速、简便、安全而可靠地检测特异mRNA表达的方法。
Semi_quantitative analysis is used for specific mRNA expression from minute quantity of cultured rabbit vascular endothelial cells.Total mRNA was abstracted from rabbit endothelial cells with QuickPrep D□R Micro mRNA Purification Kit.Using mRNA as a template,the relative content of Endothelin_1(ET_1)mRNA was analyzed by semi_quantitative reverse transcription and polymerase chain reaction(SqRT_PCR)method.Results showed that the undergraded mRNA was extracted from 10 5 cultured rabbit vascular endothelial cells,and its ratio of A 260 /A 280 was more than 1.8 .The quantity of ET_1 mRNA expression was determined with a method of SqRT_PCR.It is indicated that a rapid,simple,safe and reliable method of determining specific mRNA expression from minute quatity of cells is established.
出处
《广州中医药大学学报》
CAS
2000年第1期81-84,共4页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家中医药管理局资助!课题 (No . 95C0 35 )
关键词
基因表达
聚合酶链反应
MRNA
血管内皮细胞
GENE EXPRESSION/genetics
POLMERASE CHAIN REACTION/method
RNA,MESSENGER/isolation and purification
RABBITS
