1. Understanding the mechanism of action of the Plasmodium falciparum lariat debranching enzyme BME230 Computational Genomics Maria Daleke 03-16-05

2. RNA lariat debranching enzyme Debranches introns in lariat conformation in the intron turnover pathway Debranches introns in lariat conformation in the intron turnover pathway No experimental structure determination No experimental structure determination Develop assay for expressing protein Develop assay for expressing protein Site-directed mutagenesis for creating non- debranching enzyme Site-directed mutagenesis for creating non- debranching enzyme

3. PCR amplification Full-length DBR enzyme (1725 bp) C-terminus (899 bp) N-terminus (867 bp)

4. Cloning of C-terminal subunit C-terminus inserted in pET-24b Transformation of correct plasmids confirmed

5. Conservation N-term/C-term

6. Structural predictions