Leishmania donovani:: Characterization and expression of ORFF, a gene amplified from the LDI locus

被引：16

作者：

Ghosh, A

Madhubala, R

Myler, PJ

Stuart, KD

机构：

[1] Seattle Biomed Res Inst, Seattle, WA 98109 USA

[2] Univ Washington, Dept Pathobiol, Seattle, WA 98109 USA

[3] Jawaharlal Nehru Univ, Sch Life Sci, New Delhi 110067, India

来源：

EXPERIMENTAL PARASITOLOGY | 1999年 / 93卷 / 04期

关键词：

Leishmania donovani; Leishmania major; ORRF; LD1; gene amplification; gene expression; clamped homogenous field electrophoresis; chromosome; DL-dithiothreitol; ethylenediaminetetraacetic acid; ethylene; glycol-bis(beta-aminoethyl ether)-N; N; N'-tetraacetic acid; fetal bovine serum; kilobase pairs; kiloDaltons; minimal essential medium; drug concentration at which parasite; growth was inhibited by 50%; megabase pairs; 3-(N-morpholino)propanesulfonic; acid; open reading frame; phosphate-buffered saline; pH; 7.4; PBS+0.005% glucose; sodium dodecyl sulfate-polyacrylamide gel electrophoresis; phenylmethylsulfonyl fluroide; recombinant ORFF protein;

D O I：

10.1006/expr.1999.4453

中图分类号：

R38 [医学寄生虫学]; Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ; 100103 ;

摘要：

The LD1 locus is a 27.5-kb region of chromosome 35 that is conserved among all species of Leishmania and is amplified in several different isolates. Here, we report the genomic distribution of ORFF, a gene from the LDI region, and its expression at the RNA and protein levels in two Indian isolates of Leishmania donovani. In both of these isolates, ORFF was present as a single copy on chromosome 35. Densitometric analysis of ORFF mRNA abundance revealed relative abundance of 0.2 and 1.0 in AG83 and S-Lal, respectively. Antiserum against recombinant ORFF protein detected a protein of the predicted size (similar to 34 kDa) in both strains. The protein is most abundant in mid-log-phase promastigotes and has a nuclear localization. The ORFF protein is preferentially expressed in L. donovani amastigotes but, in contrast, is expressed at higher levels in L. major promasti gores. (C) 1999 Academic Press.

引用

页码：225 / 230

页数：6

共 25 条

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