A general approach to isolating Plasmodium falciparum genes using non-redundant oligonucleotides inferred from protein sequences of other organisms.
We have constructed a number of oligonucleotide probes and tested their utility in identifying various genes in Plasmodium falciparum. The probe sequences were based on known conserved regions of proteins from other organisms, coupled with an analysis of the codon usage of the parasite. By using long single oligonucleotides, we have successfully isolated the DHFR-TS gene, two actin genes and two tubulin genes from the K1 (Thailand) isolate of P. falciparum. We compare these single probes to multiply-redundant short oligonucleotide probes and to heterologous probes. We also present a detailed quantitative analysis of optimal probe design, and of how this approach can best be implemented as a general method of isolating plasmodial genes.
Hyde
J E
JE
Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology, U.K.
Kelly
S L
SL
Holloway
S P
SP
Snewin
V A
VA
Sims
P F
PF
D000818:Animals; D004247:DNA / Q000235:genetics*; D015342:DNA Probes; D005796:Genes; D015336:Molecular Probe Techniques / Q000379:methods; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010963:Plasmodium falciparum / Q000235:genetics*
D015342:DNA Probes; D015345:Oligonucleotide Probes; D004247:DNA