1Division of Animal Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243 122, (Uttar Pradesh), INDIA.
2Division of Biological Products, Indian Veterinary Research Institute, Izatnagar, 243 122, (Uttar Pradesh), INDIA.
A replication-deficient, recombinant adenovirus vector expressing LacZ transgene was used to analyse the effect of polybrene-mediated transduction in BHK-21 cells. The replication-defective recombinant adenovirus was produced by transfecting engineered adenoviral genome which contained E1a and E1b deletion and LacZ transgene, into adenoviral packaging cell line HEK-293. The produced virus was harvested and its titer was determined in terms of plaque forming units/ml (pfu/ml) in HEK-293 cells. To enhance adenovirus transduction efficiency and LacZ transgene expression, the virus vector was treated with polybrene and used to transduce BHK-21 cells. Transduction efficiency was monitored by observing percentage of cells expressing b-galactosidase. Polybrene with 6 μg/ml concentration in virus inoculum at 100 multiplicity of infection (MOI) helped to achieve approximately 95% transduction. In the control experiment, non-treated virus could achieve very little (2.3%) transduction. The results of the present study indicated that pre-treatment of recombinant adenovirus with polybrene enhanced its transduction efficiency and transgene expression in BHK-21 cells.
Adenovirus vector, BHK-21 cells, polybrene, transduction, transgene expression
