Molecular Cloning of the cDNA Encoding an Antifungal Protein in Gastrodia elata Bl.

Antifungal protein is the main inhibitor of fungal infection in the secondary corm of Gastrodia elata B1. was isolated and purified antifungal protein (GAFP) from the plant. Its molecular weight was about 14 kD. Polyclonal antibody against GAFP was produced. In vitro test, this antifungal protein inhibited the growth of some fungi in some crop including Gibberella zeae. cDNA was synthesized from poly (A) mRNA purified from G. elata. The cDNA was ligated into phage vector λgtll DNA and packaged in vitro and the phages were propagated on E. coli Y1090 and a λgtll expression library was constructed. A cDNA clone encoding for antifungal protein was screened out by immunoscreening of the library using the protein as a probe. The λDNA containing insert was digested by Eco RI after isolated and purified recombinants λDNA, the insert was obtained. The cDNA was 300 bp in length. The authors had isolated the cDNA clone encoding antifungal protein from G. elata.