Characterization of the genes for fructose-bisphosphate aldolase in Trypanosoma brucei

Marchand, Martine; Poliszczak, Annick; Gibson, Wendy; Wierenga, Rik K.; Opperdoes, Frederik; Michels, Paul A.M.

doi:10.1016/0166-6851(88)90121-1

Cited by 49 publications

(17 citation statements)

References 41 publications

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“…In order to circumvent this difficulty, an attempt has been made to determine stable isoenzyme and DNA marker of these clones, such as polymorphisms of DNA sequences encoding the glycolytic enzymes piruvate kynase 2 , fructose bi-phosphate aldolase 19 , glucose phosphate isomerase 18 and glyceraldehyde-3-phosphate dehydrogenase 16 for each parasite population 17 . In conclusion, we believe that additional behavioral and molecular markers are required for further characterization of the various T. cruzi clones present in the parasite stocks derived from Chagas disease patients.…”

Section: Discussionmentioning

confidence: 99%

Diversity of Trypanosoma cruzi stocks and clones derived from Chagas disease patients: I-Behavioral characterization in vitro

Lauria-Pires

Santana

Tavares

et al. 1997

Rev. Soc. Bras. Med. Trop.

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In this study, we isolated Trypanosoma cruzi from chronic Chagas heart disease and from megaesophagus patients. The parasite stock hSLU239 (heart disease) yielded clones h1 and h2, whereas stock mSLU142 (megaesophagus) yielded clones m1, m2, m3 and m4. The parasite growth kinetics, doubling time and differentiation in axenic liquid medium showed broad behavioral diversity. It was shown that a particular pattern of behavior for a parental stock could not necessarily be assigned for subsequent clones. This study indicates that i) each Chagas disease patient is infected with several T. cruzi populations; ii) clonal lines derived from patient samples may have different biological characteristics from the original isolate; and that iii) additional behavioral and/or molecular markers are required for further characterization of Trypanosoma cruzi stocks and clones derived from Chagas disease patients in order to identify correlations with pathology.
Neste estudo, foram obtidos estoques de Trypanosoma cruzi de pacientes chagásicos com a doença cardíaca ou com megaesôfago. O estoque hSLU239 (doença cardíaca) forneceu os clones h1 e h2, enquanto o estoque mSLU142 (megaesôfago) forneceu os clones m1, m2, m3 e m4. A cinética de crescimento do parasito, tempo de duplicação e diferenciação em meio líquido axênico mostraram ampla diversidade comportamental. Observou-se que um padrão particular de comportamento de um estoque parental podia não ser necessariamente encontrado na linhagem subclonal subseqüente. Este estudo indica que i) cada paciente chagásico é infectado com várias subpopulações de T. cruzi; ii) linhagens clonais derivadas de cada estoque do parasito podem ter características biológicas diferentes do isolado original de paciente chagásico; e que iii) marcadores comportamentais e/ou moleculares adicionais são necessários para melhor caracterização de estoques de T. cruzi e seus clones derivados de pacientes com doença de Chagas, a fim de identificar as possíveis correlações com a patologia

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Section: Discussionmentioning

confidence: 99%

Diversity of Trypanosoma cruzi stocks and clones derived from Chagas disease patients: I-Behavioral characterization in vitro

Lauria-Pires

Santana

Tavares

et al. 1997

Rev. Soc. Bras. Med. Trop.

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“…Gels were blotted in the standard way (Southern, 1975) and nitrocellulose filters were hybridized as previously described with the following oligo-labelled DNA (Osinga et al, 1985) ; (7) glucose phosphate isomerase (GPI), 1-9 kb HindIII-BamHI fragment from genomic clone (Marchand et al, 1989) ; (8) pyruvate kinase (PYK), 1.2 kb PstI-EcoRI fragment from genomic DNA clone pTgPK5 ; (9) cytosolic glyceraldehyde phosphate dehydrogenase (cGAPDH), 1.7 kb BamHI-PstI fragment from genomic DNA clone pTgGAPc6 ; (10) aldolase (ALD), 1.3 kb PCR-amplified insert from cDNA pTcALD7 (Marchand et al, 1988); (11) triose phosphate isomerase (TIM), 460 bp amplified fragment spanning polymorphic KpnI site within gene from genomic clone pTgTIM12 (Swinkels et al, 1986); (12) …”

Section: Methodsmentioning

confidence: 99%

Intraclonal mating in Trypanosoma brucei is associated with out-crossing

et al. 1997

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To examine whether mating can occur within as well as between clones of Trypanosoma brucei, we transformed three T. brucei subspecies stocks with heterologous genes conferring resistance to either hygromycin or Geneticin and carried out a series of inter-and intraclone matings in all possible double drug combinations. Double drug-resistant hybrids were recovered from three of the six out-crosses, but not from any of the three intraclone matings. However, further analysis of cloned progeny trypanosomes from one of the out-crosses using RFLP markers, molecular karyotyping and RAPD (random amplification of polymorphic DNA) produced unequivocal evidence that intraas well as interclone mating had occurred. The progeny of interclone mating were double drug-resistant and heterozygous at 9 of 13 loci examined. In contrast, the progeny of intraclone mating had no demonstrable input of genetic material from the hygromycin-resistant parent and were similar to the Geneticin-resistant parent for most markers, except for five loci which were heterozygous in the Geneticin-resistant parent but homozygous in these clones (aldolase, THTl glucose transporter, procyclin, tubulin and cDNA 23). In addition, PFGE showed considerable karyotypic rearrangements in these clones and loss of genetic material was evident from RAPD and VSG (variant surface glycoprotein) gene fingerprint analysis. We conclude that intraclone mating can occur in trypanosomes, but only during out-crossing, suggesting that meiosis and/or fusion are triggered by a diffusible factor.

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“…Six additional chromosome pairs could be identified, and most of these pairs were characterized on the basis of the location of two different marker genes. These chromosome pairs are bracketed at the left side of 14 in different antigenic variants. This band can be separated into two bands in some variants, presumably due to a DNA rearrangement event which affected the size of one of the chromosomes (Fig.…”

mentioning

confidence: 99%

Chromosome organization of the protozoan Trypanosoma brucei.

Gottesdiener¹,

Garcı́a-Añoveros²,

Lee³

et al. 1990

Mol. Cell. Biol.

103

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The genome of the protozoan Trypanosoma brucei is known to be diploid. Karyotype analysis has, however, failed to identify homologous chromosomes. Having refined the technique for separating trypanosome chromosomes (L. H. T. Van der Ploeg, C. L. Smith, R. I. Polvere, and K. Gottesdiener, Nucleic Acids Res. 17:3217-3227, 1989), we can now provide evidence for the presence of homologous chromosomes. By determining the chromosomal location of different genetic markers, most of the chromosomes (14, excluding the minichromosomes), could be organized into seven chromosome pairs. In most instances, the putative homologs of a pair differed in size by about 20%. Restriction enzyme analysis of chromosome-sized DNA showed that these chromosome pairs contained large stretches of homologous DNA sequences. From these data, we infer that the chromosome pairs represent homologs. The identification of homologous chromosomes gives valuable insight into the organization of the trypanosome genome, will facilitate the genetic analysis of T. brucei, and suggests the presence of haploid gametes.

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Characterization of the genes for fructose-bisphosphate aldolase in Trypanosoma brucei

Cited by 49 publications

References 41 publications

Diversity of Trypanosoma cruzi stocks and clones derived from Chagas disease patients: I-Behavioral characterization in vitro

Diversity of Trypanosoma cruzi stocks and clones derived from Chagas disease patients: I-Behavioral characterization in vitro

Intraclonal mating in Trypanosoma brucei is associated with out-crossing

Chromosome organization of the protozoan Trypanosoma brucei.

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