Cloning of cold-inducible dehydrin-like genes from the blackcurrant (Ribes nigrum L.) using RT-PCR.
A method is described for the identification of conserved genes in one plant species by using sequence information on internal motifs from well-characterized clones from another species. This sequence information is used to design primers for reverse transcriptase polymerase chain reaction (RT-PCR) and to design oligonucleotide probes to identify genuine positive amplification products. The approach was successfully used to clone cDNAs encoding cold-inducible dehydrin-like genes from the woody perennial blackcurrant, Ribes nigrum L. The strategy described can accelerate the cloning of heterologous cDNAs and is a convenient alternative to direct screening of cDNA libraries.
Kemp
R
R
Department of Biosciences, University of Hertfordshire, UK.
Brennan
R
R
Lanham
P G
PG
Slater
R
R
Jones
H
H
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003080:Cold Temperature; D005638:Fruit / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
D010940:Plant Proteins; C075590:dehydrin-like protein, plant