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新鲜分离的人呼吸道上皮细胞中基因表达的定量评估。

Quantitative evaluation of gene expression in freshly isolated human respiratory epithelial cells.

作者信息

Trapnell B C

机构信息

Department of Virology, Genetic Therapy, Gaithersburg, Maryland 20878.

出版信息

Am J Physiol. 1993 Mar;264(3 Pt 1):L199-212. doi: 10.1152/ajplung.1993.264.3.L199.

Abstract

Recent advances in molecular biology have led to development of methods to evaluate the in vivo expression of specific genes in epithelial cells of the human respiratory tract. These methods involve collection of small samples of respiratory epithelium from the human airway with a cytology brush, together with sensitive techniques to enumerate cells and to detect and quantify RNA using the polymerase chain reaction (PCR). Thus, using the typical kinds of tissue samples obtainable in a clinical setting, one can now evaluate in vivo expression of specific genes in human airway epithelium. Specifically, mRNA transcript levels can be determined on a copy number per cell basis even when the level of expression is in the range of 1-10 mRNA transcripts/cell. The currently available techniques for measuring mRNA transcript levels in small tissue samples are reviewed with emphasis on the use of quantitative PCR assays.

摘要

分子生物学的最新进展促使了一些方法的开发,用于评估人类呼吸道上皮细胞中特定基因的体内表达。这些方法包括用细胞学刷从人类气道采集呼吸道上皮的小样本,以及使用聚合酶链反应(PCR)来计数细胞、检测和定量RNA的敏感技术。因此,利用临床环境中可获得的典型组织样本类型,现在可以评估人类气道上皮中特定基因的体内表达。具体而言,即使表达水平在1-10个mRNA转录本/细胞的范围内,也可以在每个细胞的拷贝数基础上确定mRNA转录本水平。本文综述了目前用于测量小组织样本中mRNA转录本水平的技术,重点介绍了定量PCR检测方法的应用。

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