Stable DNA transfection of a wide range of trypanosomatids.

Abstract:

:We have shown that the Leishmania major transfection vector pR-NEO (or derivatives thereof) can be introduced and stably maintained in four species complexes of pathogenic Leishmania (L. tropica, L. mexicana, L. donovani, L. braziliensis), and the genera Endotrypanum and Crithidia; transfection of Trypanosoma cruzi or Trypanosoma brucei was not successful. Quantitative plating assays showed that the transfection efficiencies were high in L. major and Leishmania amazonensis (5x10(-5)/cell) and about 10-fold less for Leishmania panamaensis and Crithidia. Leishmania donovani transfected with pR-NEO retained the ability to infect hamsters, and amastigotes recovered after 2 months yielded G418-resistant promastigotes which retained high levels of extrachromosomal pR-NEO DNA. In promastigotes, the transfected DNA existed as extrachromosomal circles, and expressed the predicted 2.4-kb hybrid NEO/DHFR-TS mRNA bearing the trans-spliced miniexon. Large quantitative differences were observed only in Crithidia: relative to transfected Leishmania species, the copy number of pR-NEO was elevated 20-fold, while the levels of the NEO/DHRFR-TS mRNA or Escherichia coli beta-galactosidase (synthesized from the expression vector pX-beta GAL) were reduced 80 and more than 1000-fold, respectively. Thus, genetic signals derived from L. major DNA that mediate RNA expression or stability are recognized by the heterologous Leishmania species but less efficiently by Crithidia. These studies suggest that pR-NEO derived vectors may be applied to the study of genes expressed throughout the life cycle in a wide range of pathogenic trypanosomatids.

journal_name

Mol Biochem Parasitol

authors

Coburn CM,Otteman KM,McNeely T,Turco SJ,Beverley SM

doi

10.1016/0166-6851(91)90210-w

subject

Has Abstract

pub_date

1991-05-01 00:00:00

pages

169-79

issue

1

eissn

0166-6851

issn

1872-9428

pii

0166-6851(91)90210-W

journal_volume

46

pub_type

杂志文章
  • Isolation, sequencing and expression of the gene encoding hypoxanthine-guanine-xanthine phosphoribosyltransferase of Tritrichomonas foetus.

    abstract::We have cloned and expressed the full-length gene encoding the hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase) from the anaerobic protozoan parasite Tritrichomonas foetus. This enzyme is essential in nucleic acid metabolism of T. foetus because the parasite is unable to synthesize purine nucleotide...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(94)90058-2

    authors: Chin MS,Wang CC

    更新日期:1994-02-01 00:00:00

  • The mitochondrial phosphate carrier TbMCP11 is essential for mitochondrial function in the procyclic form of Trypanosoma brucei.

    abstract::Conserved amongst all eukaryotes is a family of mitochondrial carrier proteins (SLC25A) responsible for the import of various solutes across the inner mitochondrial membrane. We previously reported that the human parasite Trypanosoma brucei possesses 26 SLC25A proteins (TbMCPs) amongst which two, TbMCP11 and TbMCP8, w...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/j.molbiopara.2020.111275

    authors: Gao F,Voncken F,Colasante C

    更新日期:2020-05-01 00:00:00

  • Cloning, heterologous expression in Escherichia coli and characterization of a protein disulfide isomerase from Fasciola hepatica.

    abstract::A Fasciola hepatica cDNA clone of 1752 bp was isolated from an adult worm cDNA expression library by immunological screening using a rabbit serum against the excretory-secretory antigens. The nucleotide sequence of the cDNA revealed the presence of an open reading frame of 489 codons which encoded a 55 kDa polypeptide...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(02)00269-4

    authors: Salazar-Calderón M,Martín-Alonso JM,Castro AM,Parra F

    更新日期:2003-01-01 00:00:00

  • Characterization of Leishmania major phosphatidylethanolamine methyltransferases LmjPEM1 and LmjPEM2 and their inhibition by choline analogs.

    abstract::Phosphatidylcholine (PC) is the most abundant phospholipid in the membranes of the human parasite Leishmania. It is synthesized via two metabolic routes, the de novo pathway that starts with the uptake of choline, and the threefold methylation of phosphatidylethanolamine. Choline was shown to be dispensable for Leishm...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/j.molbiopara.2014.08.005

    authors: Bibis SS,Dahlstrom K,Zhu T,Zufferey R

    更新日期:2014-09-01 00:00:00

  • Crystal structures of Toxoplasma gondii pterin-4a-carbinolamine dehydratase and comparisons with mammalian and parasite orthologues.

    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

    authors: Cameron S,Fyffe SA,Goldie S,Hunter WN

    更新日期:2008-04-01 00:00:00

  • The immunodominant 17-kDa antigen from Cryptosporidium parvum is glycosylphosphatidylinositol-anchored.

    abstract::Cryptosporidium parvum is a protozoan parasite of the intestinal epithelium that has caused numerous outbreaks of diarrheal illness in humans. During our studies of the host immune response to C. parvum infection, we noted that two of the immunodominant surface antigens of the sporozoite stage of the parasite readily ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(00)00386-8

    authors: Priest JW,Xie LT,Arrowood MJ,Lammie PJ

    更新日期:2001-03-01 00:00:00

  • Upstream regulatory elements controlling expression of the Entamoeba histolytica lectin.

    abstract::Entamoeba histolytica genomic organization and putative promoter elements appear to be distinct from both metazoan and better characterized protozoan organisms. The recent development of DNA-mediated transfection for E. histolytica enabled characterization of cis-acting promoter elements required for gene expression. ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(96)02614-x

    authors: Purdy JE,Pho LT,Mann BJ,Petri WA Jr

    更新日期:1996-06-01 00:00:00

  • Three acetylcholinesterases of the pinewood nematode, Bursaphelenchus xylophilus: insights into distinct physiological functions.

    abstract::Acetylcholinesterase (AChE) plays a key role in postsynaptic transmission in most animals. Nematodes encode multiple AChEs, implying its functional diversity. To explore physiological functions of multiple AChEs, three distinct AChEs (BxACE-1, BxACE-2, and BxACE-3) were identified and characterized from the pinewood n...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/j.molbiopara.2010.11.005

    authors: Kang JS,Lee DW,Choi JY,Je YH,Koh YH,Lee SH

    更新日期:2011-02-01 00:00:00

  • Crystal structure and putative substrate identification for the Entamoeba histolytica low molecular weight tyrosine phosphatase.

    abstract::Entamoeba histolytica is a eukaryotic intestinal parasite of humans, and is endemic in developing countries. We have characterized the E. histolytica putative low molecular weight protein tyrosine phosphatase (LMW-PTP). The structure for this amebic tyrosine phosphatase was solved, showing the ligand-induced conformat...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/j.molbiopara.2014.01.003

    authors: Linford AS,Jiang NM,Edwards TE,Sherman NE,Van Voorhis WC,Stewart LJ,Myler PJ,Staker BL,Petri WA Jr

    更新日期:2014-01-01 00:00:00

  • Characterisation of protein kinase C like activities in Trypanosoma brucei.

    abstract::Protein kinase activities in bloodstream and procyclic forms of Trypanosoma brucei have been partially purified and characterised. Cytosolic extracts were separated on DEAE-cellulose and assayed for the ability to phosphorylate histone in the presence of Ca2+ and diacylglycerol. Five peaks of activity were identified ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(90)90135-9

    authors: Keith K,Hide G,Tait A

    更新日期:1990-11-01 00:00:00

  • Characterization of macromolecular transport pathways in malaria-infected erythrocytes.

    abstract::We have previously provided evidence for a pathway in Plasmodium falciparum-infected erythrocytes, coined the parasitophorous duct pathway, which provides serum (macro)molecules direct access to intraerythrocytic parasites . The present study addresses the purity of the fluorescent macromolecules used to define the du...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(97)00039-x

    authors: Goodyer ID,Pouvelle B,Schneider TG,Trelka DP,Taraschi TF

    更新日期:1997-07-01 00:00:00

  • Perturbation of sterol biosynthesis by itraconazole and ketoconazole in Leishmania mexicana mexicana infected macrophages.

    abstract::The azole antifungals ketoconazole and itraconazole possess in vitro antileishmanial activity against Leishmania mexicana mexicana amastigotes in macrophages (cell line J774G8). As in yeast and fungi, the activity is likely to be due to inhibition of the cytochrome P-450-dependent 14 alpha-demethylation of lanosterol ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(89)90026-1

    authors: Hart DT,Lauwers WJ,Willemsens G,Vanden Bossche H,Opperdoes FR

    更新日期:1989-03-01 00:00:00

  • Expression of a stage-specific lipophosphoglycan in Leishmania major amastigotes.

    abstract::Amastigotes of Leishmania major were isolated from infected mice and radiolabeled for 2 h with [3H]galactose. An acidic [3H]glycoconjugate was extracted from a dilipidated residue fraction with the solvent water/ethanol/diethylether/pyridine/NH4OH (15:15:5:1:0.017). The radioactivity labeled glycoconjugate was found t...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(91)90030-a

    authors: Turco SJ,Sacks DL

    更新日期:1991-03-01 00:00:00

  • Purification and characterization of gamma-glutamyl transpeptidase from Ascaris suum.

    abstract::gamma-Glutamyl transpeptidase, which is of central importance in the degradation of glutathione, was purified from Ascaris suum to apparent homogeneity. The enzyme was found to be a predominantly membrane-bound protein and was solubilized by Triton X-100. The purified enzyme, which exhibits a specific activity of 1009...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(96)02573-x

    authors: Hussein AS,Walter RD

    更新日期:1996-04-01 00:00:00

  • Translation initiation factors GleIF4E2 and GleIF4A can interact directly with the components of the pre-initiation complex to facilitate translation initiation in Giardia lamblia.

    abstract::Translation initiation factor eIF4F is essential for cap-dependent translation initiation in eukaryotes. eIF4F is a trimeric complex consisting of a scaffold protein eIF4G, cap-binding protein eIF4E and DEAD-box RNA helicase eIF4A. eIF4F binds to the 5' cap structure of the mRNA through eIF4E and facilitates the bindi...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/j.molbiopara.2020.111258

    authors: Adedoja AN,McMahan T,Neal JP,Hamal Dhakal S,Jois S,Romo D,Hull K,Garlapati S

    更新日期:2020-03-01 00:00:00

  • Amplification and sequencing of genomic DNA fragments encoding cysteine proteases from protozoan parasites.

    abstract::Cysteine protease gene fragments from three protozoan parasites Trypanosoma cruzi, Trypanosoma brucei, and Entamoeba histolytica were amplified by the polymerase chain reaction (PCR) from genomic DNA using degenerate oligonucleotide primers. The primers used for the amplification were designed based upon amino acid se...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(90)90002-4

    authors: Eakin AE,Bouvier J,Sakanari JA,Craik CS,McKerrow JH

    更新日期:1990-02-01 00:00:00

  • The kinetic properties and sensitivities to inhibitors of lactate dehydrogenases (LDH1 and LDH2) from Toxoplasma gondii: comparisons with pLDH from Plasmodium falciparum.

    abstract::Toxoplasma gondii differentially expresses two forms of lactate dehydrogenase in tachyzoites and bradyzoites, respectively, designated LDH1 and LDH2. Previously it was demonstrated that LDH1 and LDH2 share a unique structural feature with LDH from the malarial parasite Plasmodium falciparum (pLDH), namely, the additio...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(01)00360-7

    authors: Dando C,Schroeder ER,Hunsaker LA,Deck LM,Royer RE,Zhou X,Parmley SF,Vander Jagt DL

    更新日期:2001-11-01 00:00:00

  • Long-range restriction maps of size-variable homologous chromosomes in Leishmania infantum.

    abstract::In order to clarify the interpretation of molecular karyotype polymorphisms in Leishmania, the three smallest chromosomes from five cloned strains of Leishmania infantum were identified by chromosome-specific anonymous DNA probes. The presence in the same clone of homologous chromosomes of a different size was demonst...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(91)90053-9

    authors: Blaineau C,Bastien P,Rioux JA,Roizès G,Pagès M

    更新日期:1991-06-01 00:00:00

  • Genetic and antigenic characterization of Babesia bovis merozoite spherical body protein Bb-1.

    abstract::A Babesia bovis merozoite protein, Bb-1, was localized by immunoelectron microscopy to an apical organelle known as the spherical body. This unique structure appears to be analogous to dense granules of other apicomplexan protozoa. Similar to previously described dense granule proteins of Plasmodium spp., Bb-1 is secr...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(94)00200-7

    authors: Hines SA,Palmer GH,Brown WC,McElwain TF,Suarez CE,Vidotto O,Rice-Ficht AC

    更新日期:1995-02-01 00:00:00

  • The reversible effect of glucose on the energy metabolism of Schistosoma mansoni cercariae and schistosomula.

    abstract::This study on isolated cercarial bodies demonstrates that the biological transformation from cercaria to schistosomulum and the biochemical transition from an aerobic to an anaerobic energy metabolism are separate processes, which are not necessarily linked. The metabolic transition depends on the external glucose con...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(92)90202-u

    authors: Horemans AM,Tielens AG,van den Bergh SG

    更新日期:1992-03-01 00:00:00

  • Is point mutagenesis a mechanism for antigenic variation in Trypanosoma brucei?

    abstract::Antigenic variation in African trypanosomes proceeds by switching between different variant surface glycoprotein (VSG) molecules, whose extensive epitope differences enable evasion of antibody responses. Each trypanosome has approximately 1000 basic copy VSG genes inside chromosomes and a subset located at telomeres. ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(96)02642-4

    authors: Graham VS,Barry JD

    更新日期:1996-07-01 00:00:00

  • Molecular characterization and expression of two putative protective excretory secretory proteins of Haemonchus contortus.

    abstract::It has been shown that vaccination with two low molecular mass excretory secretory (ES) antigens of 15 and 24 kDa, respectively, afforded a substantial degree of protection against Haemonchus contortus to sheep. In vitro cultivation of the parasite usually yields a limited amount of these proteins and therefore, recom...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(97)00093-5

    authors: Schallig HD,van Leeuwen MA,Verstrepen BE,Cornelissen AW

    更新日期:1997-09-01 00:00:00

  • Theileria parva 104 kDa microneme--rhoptry protein is membrane-anchored by a non-cleaved amino-terminal signal sequence for entry into the endoplasmic reticulum.

    abstract::The 104 kDa microneme-rhoptry protein (p104) is the only known apical complex organelle-specific protein of Theileria parva. p104 exhibits striking structural similarities to circumsporozoite protein and sporozoite surface protein 2 of Plasmodium yoelii. Their primary sequences contain two hydrophobic segments, locate...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(99)00020-1

    authors: Ebel T,Gerhards J,Binder BR,Lipp J

    更新日期:1999-05-15 00:00:00

  • Characterization of the gene family encoding a host-protective antigen of the tapeworm Taenia ovis.

    abstract::Genomic structure has been determined for a gene encoding a host-protective antigen of the parasitic platyhelminth Taenia ovis. An incomplete cDNA, known as 45W, encodes the protective antigen. Southern hybridisation experiments using 45W cDNA as a probe, revealed that the 45W gene was a member of a multigene family. ...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(94)00104-u

    authors: Waterkeyn JG,Lightowlers MW,Coppel R,Cowman AF

    更新日期:1995-07-01 00:00:00

  • An evaluation of different methods for labelling the surface of the filarial nematode Brugia pahangi with 125iodine.

    abstract::The specificity of a range of 125I labelling techniques (Chloramine T, Iodogen, Bolton and Hunter reagent, lactoperoxidase and iodosulfanilic acid) to the surface of the filarial nematode Brugia pahangi was evaluated by autoradiography of sections of labelled worms and of dried SDS-polyacrylamide gels following electr...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(85)90091-x

    authors: Marshall E,Howells RE

    更新日期:1985-06-01 00:00:00

  • Expression and substrate specificity of a recombinant cysteine proteinase B of Leishmania braziliensis.

    abstract::The cysteine proteinase B of Leishmania parasites is an important virulence factor. In this study we have expressed, isolated and characterized for the first time a recombinant CPB from Leishmania braziliensis, the causative agent of mucocutaneous leishmaniosis. The mature region of the recombinant CPB shares a high p...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/j.molbiopara.2008.06.005

    authors: Lanfranco MF,Loayza-Muro R,Clark D,Núñez R,Zavaleta AI,Jimenez M,Meldal M,Coombs GH,Mottram JC,Izidoro M,Juliano MA,Juliano L,Arévalo J

    更新日期:2008-10-01 00:00:00

  • Differences in substrate specificity and kinetic properties of the recombinant hexokinases HXK1 and HXK2 from Entamoeba histolytica.

    abstract::Entamoeba histolytica is responsible for amoebic colitis and liver abscess in humans. Entamoeba dispar is a closely related, morphologically indistinguishable nonpathogenic species. The hexokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1) isoenzyme patterns distinguish the pathogenic and nonpathogenic species. B...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/s0166-6851(99)00164-4

    authors: Kroschewski H,Ortner S,Steipe B,Scheiner O,Wiedermann G,Duchêne M

    更新日期:2000-01-05 00:00:00

  • Polymorphism of the alleles of the merozoite surface antigens MSA1 and MSA2 in Plasmodium falciparum wild isolates from Colombia.

    abstract::The degree of polymorphism and the allelic distribution of 2 major Plasmodium falciparum merozoite surface antigens (MSA1 and MSA2) have been analysed in clinical isolates from Colombia. DNA was prepared directly from patients' blood and used in PCR reactions to amplify block 2 of MSA1 and the central region from MSA2...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(91)90070-m

    authors: Snewin VA,Herrera M,Sanchez G,Scherf A,Langsley G,Herrera S

    更新日期:1991-12-01 00:00:00

  • Identification and characterization of an Onchocerca volvulus cDNA clone encoding a highly immunogenic calponin-like protein.

    abstract::The identification and characterization of a recombinant cDNA clone, designated OV9M, expressing an antigen present in Onchocerca volvulus infective larvae and adult stages is described. Clone OV9M was identified by screening a lambda gt11 cDNA expression library derived from adult O. volvulus mRNA using pooled rabbit...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/0166-6851(94)90122-8

    authors: Irvine M,Huima T,Prince AM,Lustigman S

    更新日期:1994-05-01 00:00:00

  • Base J glucosyltransferase does not regulate the sequence specificity of J synthesis in trypanosomatid telomeric DNA.

    abstract::Telomeric DNA of trypanosomatids possesses a modified thymine base, called base J, that is synthesized in a two-step process; the base is hydroxylated by a thymidine hydroxylase forming hydroxymethyluracil (hmU) and a glucose moiety is then attached by the J-associated glucosyltransferase (JGT). To examine the importa...

    journal_title:Molecular and biochemical parasitology

    pub_type: 杂志文章

    doi:10.1016/j.molbiopara.2016.01.005

    authors: Bullard W,Cliffe L,Wang P,Wang Y,Sabatini R

    更新日期:2015-12-01 00:00:00