Project Number: 6202-22000-026-05
Project Type: Specific Cooperative Agreement

Start Date: Sep 08, 2004
End Date: Jul 14, 2009

Objective:
Identify and purify the protein or the enzyme that controls the biosynthesis of (+)gossypol in flower petals and obtain its partial amino acid sequence. Based on the partial amino acid sequence, clone and express the gene that codes for the protein or enzyme that controls the biosynthesis of (+)-gossypol in flower petals (+G2), and heterologously express it. Use the cDNA of the +G2 as a probe to identify +G1, the gene that controls the synthesis of (+)-gossypol in the seed.

Approach:
A crude enzyme extract will be prepared from flower petals collected from BC1S1 plants that exhibit only the +G2 gene (i.e., the gene that primarily controls (+)-gossypol synthesis in flower petals). This material will be purified by gel permeation and ion exchange chromatography. Fractions will be assayed for activity. The purified protein will be sequenced and the gene will be cloned. Its authenticity will be verified by heterologous expression. We will use the cDNA of this gene as a probe to identify the gene that controls the biosynthesis of (+)-gossypol in seed.