Construction of green fluorescent protein retroviral vector and its expression in various cell types

Aim To achieve efficient, and stable expression of green fluorescent protein(GFP) reporter gene in tissue-engineered cells. Methods GFP retrovirus expression vector was constructed and transfected into packaging cell PT67. After G418 selection and cell cloning, cell clones producing high levels of recombinant virus were obtained, The viruses were used to infect target cells directly. Results Bright green fluorescence of the transfected cells can be observed under fluorescent microscopes 48 hrs after transfection. The transfection rates were 20%～50%. The effective expression of GFP in target cells infected by recombinant retrovirus has lasted for 2 months. Conclusion The construction of recombinant retrovials GFP vector can provide a simple, sensitive and reliable tool for labeling tissue engineered cell and cell dynamic research.