Plant biotech & Risk Assessment
Nguyen, Vincent , Olave, Marcella , Cohen, Amybeth .
Expression of the Anti-cocaine Antibody Fab Fragments in Chlamydomonas reinhardtii.
Chlamydomonas reinhardtii is an attractive candidate for heterologous protein expression as it reproduces rapidly, can be cultured inexpensively in large volumes, carries out most post-translation modifications, and allows for site-specific integration of the gene of interest into all copies of the circular chloroplast genome. To date, the heavy and light chain genes of the anti-cocaine Fab fragment have been synthesized to reflect codon usage in the chloroplast of Chlamydomonas reinhardtii. Mutation-free copies of the genes, containing the light-regulated atpA or psbD promoters and rbcL terminator, were integrated into the chloroplast genome using microprojectile bombardment. Accumulation of the light chain protein, containing an M2 epitope tag, was easily detected. However, the heavy chain protein, containing the same M2 tag, accumulated to very low levels. As northern blot analysis showed that the light and heavy chain mRNAs accumulated to approximately equal levels, translation of the heavy chain mRNA may be affected. Experiments in which cells are shifted from the dark into the light for varying periods of time are under way in order to assess protein production. Analysis of polyribosomal RNA will determine if the light chain mRNA is being translated more efficiently than the heavy chain mRNA, or if protein stability is affected. The successful production of functional anti-cocaine antibodies may be utilized in drug testing and the treatment of cocaine overdose, and will further confirm Chlamydomonas reinhardtii as a viable system for the expression of therapeutic proteins.
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1 - California State University, Fullerton, Biological Science, 800 N. State College Blvd, Fullerton, CA, 92834, USA
2 - California State University, Fullerton, Biological Science
antibody genes in plants.
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM