In order to establish malaria parasite transfection technology in South Africa, firefly luciferase and green fluorescent protein (GFP) reporter constructs were prepared. In attempt to simplify these constructs, a var intron (PFC0005w), previously reported to have bidirectional promoter activity, was utilized to drive expression through two genes (i.e. the antibiotic-resistance gene, human dhfr and the reporter gene) in a head-to-head orientation. In addition, protocols were adjusted by including DNA packaging reagents to improve uptake into the parasite and by using shaking instead of stationary parasite cultures to improve the parasite proliferation and selection rate. Successfully transfected parasites were selected by the antifolate WR99210.
Reference:
Van Brummelen AC, Becker JVW et al. 2010. Establishing malaria parasite transfection technology in South Africa. 22nd South African Society for Biochemistry and Molecular Biology Congress. Bloemfontein, South Africa, 18-20 January 2010, pp 2
Van Brummelen, A., Becker, J., Mancama, D. T., & Hoppe, H. (2010). Establishing malaria parasite transfection technology in South Africa. http://hdl.handle.net/10204/3971
Van Brummelen, AC, JVW Becker, Dalubuhle T Mancama, and H Hoppe. "Establishing malaria parasite transfection technology in South Africa." (2010): http://hdl.handle.net/10204/3971
Van Brummelen A, Becker J, Mancama DT, Hoppe H, Establishing malaria parasite transfection technology in South Africa; 2010. http://hdl.handle.net/10204/3971 .
