Deletion of the gene for the membrane-bound acid phosphatase of Leishmania mexicana.
The membrane-bound acid phosphatase of Leishmania mexicana (LmxMBAP) has been shown to be a heterogeneously N-glycosylated type I transmembrane protein, which is localized predominantly in vesicular structures close to the flagellar pocket in promastigotes and amastigotes. Its expression in both life stages prompted us to analyse its function by performing deletion analysis. Both alleles of the single copy gene were sequentially replaced by resistance marker genes and the resulting deletion mutant was tested for its potential to infect Balb/c mice and peritoneal macrophages. There was no obvious difference detectable between the mutant and the wild-type. Therefore, we conclude that LmxMBAP is neither involved in the infection process nor required for amastigote survival in the infected host cell. LmxMBAP null mutant promastigotes were used to establish a system for homogeneous overexpression of LmxMBAP which will be useful to investigate protein sorting in L. mexicana.
Benzel
I
I
Max-Planck-Institut für Biologie, Abteilung Membranbiochemie, Corrensstrasse 38, D-72076 Tübingen, Germany.
Weise
F
F
Wiese
M
M
D000135:Acid Phosphatase / Q000235:genetics* / Q000378:metabolism*; D000483:Alleles; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D017353:Gene Deletion; D018390:Gene Targeting; D017125:Genes, Protozoan; D020459:Genes, rRNA; D007894:Leishmania mexicana / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology; D008264:Macrophages / Q000469:parasitology; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014774:Virulence
D008565:Membrane Proteins; D015800:Protozoan Proteins; D000135:Acid Phosphatase