Pending CRIS Projects:

3655-21000-033-04T John Ralph

ALTERING LIGNIN STRUCTURE FOR IMPROVED FIBER UTILIZATION

Objectives: 1) Purify and characterize the require enzyme; 2) Develop cDNA libraries from the host plant; 3) Use purified protein sequence to develop probes for screening libraries; 4) Use PCR to screen for gene similarity; and 5) Develop strategies for molecular manipulation of appropriate model plants to test our hypothesis.

Approach: Two approaches will be undertaken to accomplish the goal of modifying lignin polymers in plants such as trees and forages. One approach is to purify and characterize the modifying enzyme, sequence the purified enzyme, and use this sequence to screen cDNA libraries to obtain the gene for transformation work. A second approach is to generate cDNA libraries from plants expressing this appropriate enzyme activity and use sequence information from known enzymes of similar activity to develop PCR amplification strategies to obtain the appropriate gene. Candidate genes will be expressed in E. coli and tested to determine the types and level of enzyme activity present. Two approaches will be undertaken to accomplish the goal of modifying lignin polymers in plants such as trees and forages. One approach is to purify and characterize the modifying enzyme, sequence the purified enzyme, and use this sequence to screen cDNA libraries to obtain the gene for transformation work. A second approach is to generate cDNA libraries from plants expressing this appropriate enzyme activity and use sequence information from known enzymes of similar activity to develop PCR amplification strategies to obtain the appropriate gene. Candidate genes will be expressed in E. coli and tested to determine the types and level of enzyme activity present.