Wayne R. Curtis (1), Noel Goddard (2), George Church (2), Joe Chappell (3)
(1) Penn State Dept Chemical Engineering, (2) Harvard Medical School, (3) U. Kentucky, Plant & Soil Science.
As part of my recent sabbatical, a collaboration was established to initiate research on genetic engineering of (isoprene) hydrocarbon synthesis in algae. This talk will present the lessons learned from this effort in the context of reviewing general progress in algae genetic engineering. Chlamydomonas reinhardtii (Chlamy) was chosen because it is a ‘model’ eukaryotic green microalgae. A recently developed ‘high GC content’ aminoglycoside resistance gene marker provides reasonably efficient transformation; however, the >65%GC content made “PCR oligo-overlap assembly” of constructs challenging. Avian farnesyl diphosphate synthase was chosen to avoid endogenous silencing; chloroplastic transit sequences will permit comparison of diverting compartmentalized isoprene fluxes with heterologous sesquiterpene synthases.