Identification of stress-responsive genes in plants using suppression subtraction hybridization: ozone stress as an example.
Among the open-ended techniques for identifying differentially expressed genes in response to stress, the PCR-based suppression subtraction hybridization (SSH) is widely used. The popularity of this technique stems from the ease of conducting this procedure in any laboratory set up for basic molecular biology research. Further, the availability of a comprehensive kit for conducting suppression subtractions from BD Biosciences has made this technique easy to adapt and adopt to any biological system. In this chapter we describe in detail the SSH procedure and explain the subtle changes that have been incorporated to make this technique adaptable for identifying stress-responsive genes in plants.
Peal
Lila
L
Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK, USA.
Puckette
Michael
M
Mahalingam
Ramamurthy
R
D017360:Arabidopsis / Q000187:drug effects* / Q000235:genetics*; D018076:DNA, Complementary / Q000096:biosynthesis; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D015723:Gene Library; D017343:Genes, Plant / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010126:Ozone / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000302:isolation & purification; D018749:RNA, Plant / Q000302:isolation & purification; D013312:Stress, Physiological / Q000187:drug effects / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D018749:RNA, Plant; D010126:Ozone; D015252:Deoxyribonucleases, Type II Site-Specific; C040158:GTAC-specific type II deoxyribonucleases