Transfection of the malaria parasite and expression of firefly luciferase.
The goal of this work is to develop a method for the functional analysis of malaria genes using the method of DNA transfection. We have developed a transient transfection vector by constructing a chimeric gene in which the firefly luciferase gene was inserted in frame into the coding region of the pgs28 gene of Plasmodium gallinaceum. This plasmid DNA was introduced into P. gallinaceum gametes and fertilized zygotes by electroporation, and luciferase expression was assayed after 24 hr. This report of successful introduction and expression of a foreign gene in a malaria parasite demonstrates the feasibility of this approach to developing methods for the functional analysis of parasite genes.
Department of Tropical Public Health, Harvard School of Public Health, Boston, MA 02115.
R
R
Goonewardene
J
J
Daily
D
D
Kaslow
T J
TJ
Sullivan
P
P
Duffy
R
R
Carter
K
K
Mendis
D
D
Wirth
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D002678:Chimera; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008289:Malaria, Avian / Q000097:blood; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D010964:Plasmodium gallinaceum / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D014162:Transfection
D016054:DNA, Protozoan; D009838:Oligodeoxyribonucleotides; D008156:Luciferases