Competitive hybridization: theory and application in isolation and quantification of differentially regulated genes.
Competitive hybridization is a simple yet powerful method that was developed to screen cDNA libraries for differentially regulated genes. The method is based on competition between unlabeled cDNA from the mRNA of one sample and labeled cDNA from another sample. By manipulating the amount of competing unlabeled cDNA, background signals from the nonregulated genes can be increased or reduced, enabling the signals from differentially regulated genes to be contrasted and to be identified in a quantitative manner. To demonstrate the feasibility of the method, we screened a citrus cDNA library for ethylene-induced genes and identified three genes with different levels of ethylene induction. The mathematical basis of the method and its possible application in gene chip technology are discussed.
Zhong
G Y
GY
Kenedy-Leigh Centre for Horticultural Research, Hebrew University of Jerusalem, Rehovot, 76-100, Israel.
Riov
J
J
Goren
R
R
Holland
D
D
D015152:Blotting, Northern; D015139:Blotting, Southern; D002957:Citrus / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000032:analysis; D005030:Ethylenes / Q000494:pharmacology; D005786:Gene Expression Regulation; D015723:Gene Library; D007700:Kinetics; D008962:Models, Theoretical; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010957:Plasmids / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism
D018076:DNA, Complementary; D018744:DNA, Plant; D005030:Ethylenes; D012333:RNA, Messenger; C036216:ethylene