Isolation and characterization of a cDNA corresponding to a stress-activated cyclophilin gene in Solanum commersonii

Abstract

(Dynal, Norway). Solid phase reverse transcription of both A cDNA clone encoding cyclophilin (CyP), was isolated from poly-A+ RNA pools was performed to prepare immobilized Solanum commersonii by utilizing subtractive hybridization. sscDNA libraries. The immobilized sscDNAs from target cells The corresponding gene was found to be constitutively was used as a template in second strand cDNA synthesis and expressed in S. commersonii leaves. However, when the 32P-dCTP was added to the reaction for labelling of the syntheplants were exposed to low temperature, abscisic acid (ABA), sized cDNA fragments. After synthesis the cDNA strands drought, and wounding the amount of the cyclophilin mRNA were separated and the templates collected by a magnet. The was markedly increased. In addition, the gene was shown to labelled second strand fragments were subtracted three times be responsive to salicylic acid and pathogen challenge, sug- with the immobilized sscDNA library prepared from the subgesting a role in several different stress responses in plants. tractor cells. The remaining second strand cDNA fragments were used as a probe to screen a l-ZAP (Stratagene, USA)