Abstract：
:The technique of RNA interference (RNAi) is exceedingly useful for knocking down the expression of a specific mRNA in African trypanosomes and other organisms for the purpose of examining the function of its gene. However, when we attempted to apply RNAi in the Latin American trypanosome, Trypanosoma cruzi, to diminish expression of mRNA encoding the surface protein amastin, we found that the amastin double-stranded RNA (dsRNA) was not efficiently degraded in either epimastigotes or amastigotes, and the level of amastin mRNA remained unchanged. We generated a strain of T. cruzi CL-Brener in which the T7 promoter and tetracycline operator could be used to maximize tetracycline-regulated dsRNA synthesis and constructed plasmids that direct dsRNA against four different T. cruzi endogenous genes (encoding beta-tubulin, GP72 (flagellar adhesion protein), ribosomal protein P0 and amastin) and an exogenously added gene (GFP; green fluorescent protein). After either stable or transient transfection of these plasmids into T. cruzi, the expected RNAi phenotype was not observed for any of the five genes, although the T. cruzi beta-tubulin RNAi plasmid did give the expected FAT cell phenotype in the African trypanosome, Trypanosoma brucei. These data indicate that, similar to Leishmania, T. cruzi lacks one or more components necessary for the RNAi pathway and that these components will need to be engineered into T. cruzi, or compensated for, before RNAi can be used to study gene function in this organism.
journal_name
Mol Biochem Parasitoljournal_title
Molecular and biochemical parasitologyauthors
DaRocha WD,Otsu K,Teixeira SM,Donelson JEdoi
10.1016/j.molbiopara.2003.10.005subject
Has Abstractpub_date
2004-02-01 00:00:00pages
175-86issue
2eissn
0166-6851issn
1872-9428pii
S0166685103003025journal_volume
133pub_type
杂志文章abstract：:In trypanosomes, trans-splicing is a major essential RNA-processing mechanism that involves the addition of a spliced leader sequence to all mRNAs from a small RNA species, known as the spliced leader RNA (SL RNA). SL RNA maturation is poorly understood and it is not clear where assembly with Sm proteins takes place. ...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2006.07.004
更新日期：2006-12-01 00:00:00
abstract：:Peptidomimetic inhibitors of mammalian zinc metalloproteases have been tested as potential agents for intervention in disease caused by kinetoplastid protozoa. Certain metalloprotease inhibitors were able to inhibit the release of variant surface glycoprotein from cultured transgenic procyclic Trypanosoma brucei, conf...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(01)00244-4
更新日期：2001-04-25 00:00:00
abstract：:Two proteins were previously described in the excretory-secretory products (ES) collected from Dirofilaria immitis during the molt from the third stage to the fourth stage in vitro. The two proteins were purified using cation exchange and reverse phase HPLC. During the purification of these two proteins, a third prote...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(95)02533-2
更新日期：1996-01-01 00:00:00
abstract：:Microsporidia are highly adapted eukaryotic intracellular parasites that infect a variety of animals. Microsporidia contain no recognisable mitochondrion, but recently have been shown to have evolved from fungi and to possess heat shock protein genes derived from mitochondria. These findings make it clear that microsp...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(01)00356-5
更新日期：2001-10-01 00:00:00
abstract：:Plasmodium falciparum invades human erythrocytes by redundant pathways. Unlike Plasmodium vivax that has one Duffy Binding-Like (DBL) receptor, P. falciparum has four members of the DBL receptor family. Furthermore, one of these DBL genes, BAEBL, has polymorphisms at four amino acids in region II; each polymorphism bi...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2009.06.006
更新日期：2009-11-01 00:00:00
abstract：:The nucleotide sequence of two non-contiguous DNA fragments of 4.0 and 2.2 kb, respectively, of the kinetoplast maxicircle of Trypanosoma brucei brucei EATRO strain 427 has been determined, completing the sequence analysis of the so-called variable region (see also de Vries et al., 1988, Mol. Biochem. Parasitol. 27, 7...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(92)90178-m
更新日期：1992-12-01 00:00:00
abstract：:We have determined the sequence of a cDNA (Tt34c1) encoding a Trypanosoma cruzi trypomastigote stage-specific 85-kDa surface glycoprotein (gp85). Within the peptide sequence of Tt34c1 are two 8-amino acid motifs, Ser-X-Asp-X-Gly-X-Thr-Trp, that are characteristic of bacterial neuraminidases. Analysis of the Tt34c1 seq...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(91)90114-l
更新日期：1991-10-01 00:00:00
abstract：:The aim of this study was to investigate the occurrence, distribution and immunochemical properties of antibody-defined carbohydrate epitopes in neutral glycolipid fractions of Schistosoma mansoni eggs, cercariae and adults. The amount of extractable, antigenic, neutral glycolipids was lowest in adult worms, increasin...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(99)00123-1
更新日期：1999-10-15 00:00:00
abstract：:The enzyme pterin-4a-carbinolamine dehydratase (PCD) is important for the recycling of pterins within eukaryotic cells. A recombinant expression system for PCD from the apicomplexan parasite Toxoplasma gondii has been prepared, the protein purified and crystallised. Single crystal X-ray diffraction methods have produc...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2007.12.002
更新日期：2008-04-01 00:00:00
abstract：:A gene which overexpresses a 36-kDa protein (p36) in tunicamycin-resistant Leishmania was mapped by transfection and overexpression to the upstream region of the drug maker in the extrachromosomal amplicon. Complete sequencing of this region revealed a single open reading frame of about 1 kb. Authenticity of the clone...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(94)90147-3
更新日期：1994-08-01 00:00:00
abstract：:We have identified a GM2-activator protein (GM2AP) with highly unusual properties secreted by the nematode parasite Trichinella spiralis. Expression in Pichia pastoris resulted in a hyperglycosylated protein of 28 kDa, but the 18 kDa native protein was not glycosylated. The parasite GM2AP does not facilitate degradati...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2006.02.014
更新日期：2006-06-01 00:00:00
abstract：:Trophozoites of Entamoeba invadens were able to ingest glucopolysaccharides and metabolize them. An activity capable of degrading these substrates was purified to apparent homogeneity. The enzyme was identified as beta-amylase (alpha-1,4-D-glucan maltohydrolase EC 3.2.1.2): It was active against glycogen, amylose and ...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(96)02765-x
更新日期：1996-12-20 00:00:00
abstract：:Laminin is a major constituent of the basal lamina surrounding the midgut of the malaria vectors that has been implicated in the development of the Plasmodium oocyst. In this report we describe the cloning of the Anopheles gambiae gene encoding the laminin gamma 1 polypeptide and follow its expression during mosquito ...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(00)00371-6
更新日期：2001-02-01 00:00:00
abstract：:Thirteen monoclonal antibodies (MAbs) specific for the membrane of live Trypanosoma cruzi have been obtained from BALB/c infected mice. Most of them had greater avidity for intact than for disrupted parasites. According to the staining by indirect immunofluorescence of the different live developmental stages of the pa...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(88)90073-4
更新日期：1988-06-01 00:00:00
abstract：:The Plasmodium falciparum P-glycoprotein homologue 1 (PGH1) is structurally similar to several members of the ATP-binding cassette (ABC) superfamily of membrane transporters. We have examined whether the nucleotide binding domains predicted from the deduced amino sequence are functional by photoaffinity labeling of pu...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(93)90048-3
更新日期：1993-04-01 00:00:00
abstract：:In silico analyses of Leishmania spp. genome data are a powerful resource to improve the understanding of these pathogens' biology. Trypanosomatids such as Leishmania spp. have their protein-coding genes grouped in long polycistronic units of functionally unrelated genes. The control of gene expression happens by a va...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2012.02.009
更新日期：2012-06-01 00:00:00
abstract：:Theileria annulata is a tick-transmitted protozoan that causes tropical theileriosis, an often fatal leukoproliferative disorder of cattle. To characterize and identify parasite proteins suitable as diagnostic antigens and/or vaccine candidates, a cDNA clone encoding a macroschizont stage protein was isolated and char...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(02)00013-0
更新日期：2002-04-09 00:00:00
abstract：:The (Ca(2+)-Mg2+)ATPase activity in microsomes of Schistosoma mansoni is fully inhibited by vanadate (I50 = 2.5 microM). 45Ca2+ is accumulated within microsomal vesicles in an ATP-dependent process that is enhanced 5-fold in the presence of 40 mM phosphate. Accumulated 45Ca2+ is rapidly released by 5 microM of the Ca2...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(92)90049-p
更新日期：1992-06-01 00:00:00
abstract：:In trypanosomatids, removal of hydrogen peroxide and other aryl and alkyl peroxides is achieved by the NADPH-dependent trypanothione peroxidase system, whose components are trypanothione reductase (TRYR), trypanothione, tryparedoxin (TRYX) and tryparedoxin peroxidase (TRYP). Here, we report the cloning of a multi-copy...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/s0166-6851(01)00320-6
更新日期：2001-09-03 00:00:00
abstract：:This study further explores the effects of hypoxia and acute osmotic stress on intermediary metabolism of Leishmania major and Leishmania donovani. Late log phase promastigotes were washed and incubated with glucose as the sole exogenous carbon source, and rates of glucose consumption and product formation were measur...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(92)90217-8
更新日期：1992-02-01 00:00:00
abstract：:Plasmodium falciparum malaria parasites invade human red blood cells and immediately begin making significant alterations to the structure of the erythrocyte. These alterations facilitate the movement of nutrients into, and waste products and parasite-derived proteins out of the cell to meet the needs of the growing p...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章,评审
doi：10.1016/0166-6851(95)02575-8
更新日期：1996-02-01 00:00:00
abstract：:Cryptosporidiosis is an obligate intracellular pathogen causing diarrhea. Merozoite egress is essential for infection to spread between host cells. However, the mechanisms of egress have yet to be defined. We hypothesized that Cyclic GMP-Dependent Protein Kinase G (PKG) may be involved in Cryptosporidium egress. In th...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2020.111277
更新日期：2020-05-01 00:00:00
abstract：:Theileria annulata is an important pathogen of cattle in the tropics. The gene sequence of a sporozoite surface antigen (SPAG-1) is reported. Data is also presented demonstrating that SPAG-1 is synthesised as a large precursor. This antigen, which is a candidate for inclusion in a subunit vaccine, shows a remarkable d...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(92)90012-9
更新日期：1992-07-01 00:00:00
abstract：:The merozoite surface protein-1 of the human malaria parasite Plasmodium falciparum undergoes an extracellular proteolytic cleavage (secondary processing) intrinsic to successful erythrocyte invasion. In the T9/96 clone of P. falciparum the protease responsible has been characterised as a membrane-associated, calcium-...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(93)90182-w
更新日期：1993-11-01 00:00:00
abstract：:The glycine cleavage complex (GCV) is a potential source of the one carbon donor 5,10-methylene-tetrahydrofolate (5,10-CH(2)-THF) in the malaria parasite Plasmodium falciparum. One carbon (C1) donor units are necessary for amino acid and nucleotide biosynthesis, and for the initiation of mitochondrial and plastid tran...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2010.04.005
更新日期：2010-08-01 00:00:00
abstract：:A method for a 50-60-fold purification of a cysteine proteinase from trophozoites of Entamoeba histolytica using 35-80% ammonium sulphate fractionation, gel chromatography on Sephadex G-75, and preparative isoelectric focusing is described. The enzyme was examined for its proteolytic potencies towards native enzyme su...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(86)90054-x
更新日期：1986-01-01 00:00:00
abstract：:The capacity for rapid localization of epitope-tagged or fluorescent fusion proteins in cells is an important tool for biological discovery and functional analysis. For Trypanosoma cruzi, the protozoan parasite that causes human Chagas disease, visualization of ectopically-expressed proteins in the clinically-relevant...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2015.02.001
更新日期：2014-12-01 00:00:00
abstract：:Trypanosoma brucei harbors a unique multifunctional RNA polymerase (pol) I which transcribes, in addition to ribosomal RNA genes, the gene units encoding the major cell surface antigens variant surface glycoprotein and procyclin. In consequence, this RNA pol I is recruited to three structurally different types of prom...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/j.molbiopara.2006.02.023
更新日期：2006-09-01 00:00:00
abstract：:Giardia duodenalis is a widespread parasite of mammalian species, including humans. Due to its invariant morphology, investigation on aspects such as host specificity and transmission patterns requires a direct genetic characterization of cysts/trophozoites from host samples. A number of molecular assays have been dev...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章,评审
doi：10.1016/j.molbiopara.2008.04.006
更新日期：2008-08-01 00:00:00
abstract：:The nucleotide sequence of a gene encoding a 35-kDa thiol protease of the parasitic nematode Haemonchus contortus has been determined. The gene, designated AC-2, shares 97% nucleotide sequence identity and 98% amino acid identity with previously characterized AC-1 cDNAs encoding the thiol protease. The AC-2 gene spans...
journal_title：Molecular and biochemical parasitology
pub_type： 杂志文章
doi：10.1016/0166-6851(90)90143-a
更新日期：1990-12-01 00:00:00
