CDNA encoding a polypeptide including a hevein sequence
AssigneeBoard of Trustees operating Michigan State University
InventorsNatasha V. Raikhel, Willem F. Broekaert, Nam-Hai Chua, Anil Kush
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AssigneeBoard of Trustees operating Michigan State University
InventorsNatasha V. Raikhel, Willem F. Broekaert, Nam-Hai Chua, Anil Kush
Estimating the term…
A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli.
A fusion protein wherein maltose binding protein encoded by plasmid pIH821 is fused to a protein selected from the group consisting of the sequence of 204 amino acids shown in FIG. 2 and subfragments of said sequence larger than the 43 amino acid hevein sequence which includes the hevein sequence which binds chitin.
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