Authors : Arcelia Alvarado Islas, Angel Horacio Sandoval Trujillo, Victor R. Tenorio Gutierrez, Rogelio Alonso Morales, Octavio de Paz Villafan and Alvaro Aguilar Setien
Abstract: In previous studies with a lectin from crotalid poison that was called Fraction AL27, its capacity to penetrate the cell was detected. It was considered that such an effect made fraction AL27 usable as a facilitating agent in the cell capture of plasmids transporting genes of interest. Therefore, it was evaluated if crotalid fraction AL27 by means of the pGFP reporter plasmid could favor transfection and plasmid expression in vitro and in vivo conditions. When, carrying out transfections under the calcium/phosphate system in CHO cells and observation by Confocal Microscope (CM), a higher level of fluorescence was detected in monostrata transfected with the pGFP/AL27 mix when compared to those inoculated with the pGFP alone. By Flow Cytometry (FC) quantification, transfection efficiency was 14% for the pGFP plasmid and 24.87% for the pGFP/AL27 mix. When carrying out transfections by inoculation of the tibial muscle of mice and observation by CM, it was detected that doses of 50, 100 and 150 μg of pGFP generated a good transfection level. The minimum 50 μg of DNA doses was selected and it was observed that when 1.062 μg mL-1 of AL27 was added, fluorescence was as intense or similar to that provided by the doses of 150 μg of pGFP alone.
Arcelia Alvarado Islas, Angel Horacio Sandoval Trujillo, Victor R. Tenorio Gutierrez, Rogelio Alonso Morales, Octavio de Paz Villafan and Alvaro Aguilar Setien, 2010. Increased Expression of a Reporter Plasmid by Crotalid Venom Fraction. Journal of Animal and Veterinary Advances, 9: 2214-2219.