Selective amplification via biotin- and restriction-mediated enrichment (SABRE), a novel selective amplification procedure for detection of differentially expressed mRNAs

被引:44
作者
Lavery, DJ [1 ]
LopezMolina, L [1 ]
FleuryOlela, F [1 ]
Schibler, U [1 ]
机构
[1] UNIV GENEVA, DEPT MOL BIOL, CH-1211 GENEVA 4, SWITZERLAND
关键词
circadian expression; competitive hybridization; coumarin; 7-hydroxylase; magnetic separation; streptavidin-biotin affinity;
D O I
10.1073/pnas.94.13.6831
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We present a novel subtractive enrichment protocol for the identification of differentially expressed mRNA species. This procedure, called SABRE (selective amplification via biotin- and restriction-mediated enrichment), uses selective streptavidin-biotin affinity and restriction enzyme site reconstitution to enrich for cDNA species more abundant in one population than in another, Analysis of liver cDNA from a mouse strain expressing the neomycin resistance gene demonstrated that this procedure is capable of identifying species present in one population but absent from another. Furthermore, experiments to identify genes with circadian expression patterns in mouse liver demonstrated that SABRE is capable of detecting even modest 2- to 10-fold differences in accumulation of moderately rare mRNA species, representing as little as 0.03% of total mRNA. These experiments identified the gene encoding coumarin 7-hydroxylase as displaying circadian expression in mouse liver.
引用
收藏
页码:6831 / 6836
页数:6
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