Transient expression of luciferase in Entamoeba histolytica driven by the ferredoxin gene 5' and 3' regions.
We have successfully transfected Entamoeba histolytica trophozoites with constructs containing ferredoxin sequences fused to the reporter gene luciferase. We have determined the conditions and parameters necessary to maximise transient luciferase expression in our system. Our optimal construct gave values of 268 x 10(3) relative light units per second (RLU s-1) when assayed representing a stimulation of 18,000-fold over the control vector. Comparison of differing constructs allowed us to conclude that the 5' and 3' ferredoxin sequences are both necessary for optimal luciferase expression from our vectors. Transcriptional initiation occurs within the consensus sequence ATTCA in both construct and chromosomal ferredoxin promoters.
Gilchrist
C A
CA
Biology Department, University of York, UK.
Streets
H L
HL
Ackers
J P
JP
Hall
R
R
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity; D004749:Entamoebiasis / Q000209:etiology; D005288:Ferredoxins / Q000235:genetics*; D015870:Gene Expression; D017125:Genes, Protozoan; D005809:Genes, Regulator; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D014162:Transfection; D014774:Virulence / Q000235:genetics
D017931:DNA Primers; D016054:DNA, Protozoan; D005288:Ferredoxins; D008156:Luciferases