一种灵活的方法来研究稳定转染的哺乳动物细胞中转录后基因调控。
A flexible approach to studying post-transcriptional gene regulation in stably transfected mammalian cells.
机构信息
Veterans Administration Research Service, Veterans Affairs Medical Center, Mailstop 151, 215 North Main Street, White River Junction, VT 05009-0001, USA.
出版信息
Mol Biotechnol. 2011 Jul;48(3):210-7. doi: 10.1007/s12033-010-9360-8.
Abstract
The study of post-transcriptional regulation is constrained by the technical limitations associated with both transient and stable transfection of chimeric reporter plasmids examining the activity of 3'-UTR cis-acting elements. We report the adaptation of a commercially available system that enables consistent stable integration of chimeric reporter cDNA into a single genomic site in which transcription is induced by tetracycline. Using this system, we demonstrate the tight control afforded by this system and its suitability in mapping the regulatory function of defined cis-acting elements in the human TNF 3'-UTR, as well as the distinct effects of serum starvation on transiently transfected and stably integrated chimeric reporter genes.
摘要
该研究的转录后调控受到技术限制的约束，这与瞬时和稳定转染嵌合报告质粒来研究 3'-UTR 顺式作用元件的活性有关。我们报告了一种商业上可用的系统的适应性，该系统能够使嵌合报告 cDNA 稳定整合到单个基因组位点，四环素诱导转录。使用该系统，我们证明了该系统提供的严格控制及其在映射人 TNF 3'-UTR 中定义的顺式作用元件的调节功能方面的适用性，以及血清饥饿对瞬时转染和稳定整合嵌合报告基因的不同影响。
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