Papadopoulou B, Roy G, Dey S, Rosen B P, Olivier M, Ouellette M
Service d'Infectiologie du Centre de Recherche du CHUL, Faculté de Médecine, Université Laval, Québec, Canada.
Biochem Biophys Res Commun. 1996 Jul 25;224(3):772-8. doi: 10.1006/bbrc.1996.1098.
Transfection of pgpA into Leishmania confers resistance to arsenite and antimonials. By gene targeting mediated by homologous recombination the two alleles of the pgpA gene of a L. tarentolae wild-type cell were disrupted sequentially with the neomycin and hygromycin phosphotransferase genes. This pgpA null mutant showed an increased sensitivity to arsenite and antimonite. In addition, the L. tarentolae pgpA null mutant exhibited a decreased intracellular survival inside murine macrophages. The observed phenotypes were reverted to levels not statistically different than wild-type when an intact pgpA gene was introduced into the null mutant. Disruption of the pgpA chromosomal locus in an arsenite resistant mutant indicated that PgpA is not essential for resistance to oxyanions, although it might be required in the early steps of selection when resistance is being established.
将pgpA转染到利什曼原虫中可使其对亚砷酸盐和锑剂产生抗性。通过同源重组介导的基因靶向，依次用新霉素和潮霉素磷酸转移酶基因破坏了野生型塔兰托拉热利什曼原虫细胞pgpA基因的两个等位基因。这个pgpA基因缺失突变体对亚砷酸盐和亚锑酸盐的敏感性增加。此外，塔兰托拉热利什曼原虫pgpA基因缺失突变体在小鼠巨噬细胞内的细胞内存活率降低。当将完整的pgpA基因导入该基因缺失突变体时，观察到的表型恢复到与野生型无统计学差异的水平。在一个抗亚砷酸盐突变体中破坏pgpA染色体位点表明，PgpA对于抗氧阴离子不是必需的，尽管在建立抗性的早期选择步骤中可能需要它。
