Cloning and characterization of a thermal hysteresis (antifreeze) protein with DNA-binding activity from winter bittersweet nightshade, Solanum dulcamara
Abstract
The gene for a thermal hysteresis (antifreeze) protein (sthp-64) from the bittersweet nightshade, Solanum dulcamara, was cloned and characterized. An expression cDNA library prepared from November S. dulcamara was screened using a polyclonal antibody generated against a previously purified 67 kDa thermal hysteresis protein, and positive clones were identified and sequenced. The full-length thermal hysteresis protein gene was cloned into an Escherichia coli expression vector and expressed as a fusion protein. The putative thermal hysteresis protein (STHP-64) contains two conserved regions 56 and 57 amino acids in length which have the C-X4-C-X22-23-H-X1-H zinc finger motif which is present in WRKY proteins, a family of transcription factors which play a role in regulating expression of pathogenesis-related proteins in plants. Additional features of transcription factors, such as an acidic domain between the two zinc-fingers and a glutamine-rich region upstream of the first zinc-finger are also present in STHP-64. A DNA binding assay showed that the expressed STHP-64 fusion protein has specific DNA-binding ability. A unique feature of STHP-64 is that the C-terminus contains 10 consecutive 13-mer repeats. Such repeats are a common...Continue Reading
Citations
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