Mapping the genetic locus implicated in cytoadherence of Plasmodium falciparum to melanoma cells.
Many lines of Plasmodium falciparum undergo a deletion of the right end of chromosome 9 during in vitro cultivation accompanied by loss of cytoadherence to melanoma cells. The deletion also results in loss of expression of PfEMP1, the putative cytoadherence ligand, suggesting that PfEMP1 or a regulatory gene controlling PfEMP1 expression is encoded in this region. Initially a library of short fragments highly enriched for the right arm of chromosome 9 was constructed in bacteriophage lambda. Clones from this library were obtained randomly by the polymerase chain reaction (PCR) technique, sequenced and used to screen a yeast artificial chromosome (YAC)-P. falciparum library by PCR so that the region could be cloned and physically mapped in detail. We have used probes from this region to demonstrate that clones derived from ITG2 have undergone a deletion of intermediate length on chromosome 9. This could explain the unusual stability of cytoadherence in these clones.
Barnes
D A
DA
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Thompson
J
J
Triglia
T
T
Day
K
K
Kemp
D J
DJ
D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion / Q000235:genetics; D002874:Chromosome Mapping; D018244:Chromosomes, Artificial, Yeast; D003001:Cloning, Molecular; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D017353:Gene Deletion; D008545:Melanoma / Q000378:metabolism*; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D016054:DNA, Protozoan