| Autor: |
Souza VV; Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brasil., Vitale PAM; Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brasil., Florenzano FH; Departamento de Engenharia de Materiais, Escola de Engenharia de Lorena, Universidade de São Paulo, Lorena, SP, Brasil., Salinas RK; Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brasil., Cuccovia IM; Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brasil. |
| Abstrakt: |
Amphiphilic copolymers have a wide variety of medical and biotechnological applications, including DNA transfection in eukaryotic cells. Still, no polymer-primed transfection of prokaryotic cells has been described. The reversible addition-fragmentation chain transfer (RAFT) polymer synthesis technique and the reversible deactivation radical polymerization variants allow the design of polymers with well-controlled molar mass, morphology, and hydrophilicity/hydrophobicity ratios. RAFT was used to synthesize two amphiphilic copolymers containing different ratios of the amphiphilic poly[2-(dimethyl-amino) ethyl methacrylate] and the hydrophobic poly [methyl methacrylate]. These copolymers bound to pUC-19 DNA and successfully transfected non-competent Escherichia coli DH5α, with transformation efficiency in the range of 103 colony-forming units per µg of plasmid DNA. These results demonstrate prokaryote transformation using polymers with controlled amphiphilic/hydrophobic ratios. |
