Heat-shock tagging: a simple method for expression and isolation of plant genome DNA flanked by T-DNA insertions.
This paper describes expression profiles of the Arabidopsis HSP18.2 heat-shock gene promoter by using three different reporter genes, and the application of this promoter to a method we have developed to drive heat-shock-dependent transcription of plant genome DNA flanked by T-DNA insertions. We show that, irrespective of the location of the T-DNA insertion, an HSP18.2 promoter towards the left border of the T-DNA effectively induces transcription of flanking genome sequences in response to heat shock. If polyadenylated, tagged transcripts can be easily identified by RT-PCR.
Matsuhara
S
S
Division of Biological Sciences, Graduate School of Science, Hokkaido University, N10, W8, Sapporo 060-0810, Japan.
Jingu
F
F
Takahashi
T
T
Komeda
Y
Y
D017360:Arabidopsis / Q000235:genetics; D019677:Artificial Gene Fusion; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics*; D017930:Genes, Reporter; D018745:Genome, Plant; D006360:Heat-Shock Proteins / Q000235:genetics; D006358:Hot Temperature; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D018744:DNA, Plant; D006360:Heat-Shock Proteins; D010940:Plant Proteins; C036483:T-DNA