| Products/Services Used | Details | Operation |
|---|---|---|
| PCR Cloning and Subcloning | MATERIALS AND METHODS Expression Vector Design and Cloning The native human BChE (NCBI NM_000055) coding sequence without the native secretion signal peptide was modified for expression in rice and inserted into a vector (pUC57) containing the RAmy3D promoter, signal peptide, and terminator sequences using GenScript (GenScript, Piscataway, NJ). | Get A Quote |
An active and tetrameric form of recombinant butyrylcholinesterase (BChE), a large and complex human enzyme, was produced via semicontinuous operation in a transgenic rice cell suspension culture. After transformation of rice callus and screening of transformants, the cultures were scaled up from culture flask to a lab scale bioreactor. The bioreactor was operated through two phases each of growth and expression. The cells were able to produce BChE during both expression phases, with a maximum yield of 1.6 mg BChE/L of culture during the second expression phase. Cells successfully regrew during a 5-day growth phase. A combination of activity assays and Western blot analysis indicated production of an active and... More
