A Flexible Approach to Studying Post-Transcriptional Gene Regulation in Stably Transfected Mammalian Cells

被引：5
作者：
Nichols, Ralph C. [1 ,2 ]
Botson, John [2 ]
Wang, Xiao Wei
Hamilton, B. JoNell
Collins, Jane E.
Uribe, Victoria
Brooks, Seth A. [1 ,2 ]
Zan, Moe [2 ]
Rigby, William F. C. [2 ]
机构：
[1] Vet Affairs Med Ctr, Vet Adm Res Serv, White River Jct, VT 05009 USA
[2] Dartmouth Med Sch, Dept Med, Lebanon, NH 03756 USA
来源：
MOLECULAR BIOTECHNOLOGY | 2011年 / 48卷 / 03期
基金：
美国国家卫生研究院;
关键词：
ARE; TNF; mRNA turnover; Translation; 3 '-UTR; ACTIVATED PROTEIN-KINASE; MESSENGER-RNA DEGRADATION; TNF-ALPHA; LIPOPOLYSACCHARIDE STIMULATION; TRISTETRAPROLIN TTP; TRANSCRIPTION; EXPRESSION; TRANSLATION; BIOSYNTHESIS; INDUCTION;
D O I：
10.1007/s12033-010-9360-8
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：210 / 217
页数：8
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