A challenge to the development of this approach has been the low efficiency of standard transfection protocols particularly in primary cultures. With this approach we have shown that gfp-myosin can functionally replace the endogenous embryonic muscle myosin in C2C12 myotubes. The gfp-myosin has been isolated from infected myotubes and assayed in vitro. The development of the adenovirus gene delivery system accelerates the production of recombinant striated muscle myosin for functional assays and increases the overall yield of recombinant protein.
Recombinant adenovirus provides a versatile system for gene expression studies.
A recombinant adenoviral plasmid is rapidly generated using homologous recombination and antibiotic selection in bacteria. Homogeneous viruses are obtained from this procedure without plaque purification significantly accelerating the process of generating and testing recombinant adenoviruses. L.T. Da Costa, J. Yu ,K.W. Kinzler, and B. Vogelstein. A simplified system for generating recombinant adenoviruses. The GFP-mLC3 fluorescence marks the periodic distribution of the myosin II filaments along actin microfilaments.