Analysis of genetic complementation by whole-cell microtechniques in fibroblast heterokaryons.

Gravel, Roy A.; Leung, Anna Oi Wah; Saunders, Mary; Hösli, P.

doi:10.1073/pnas.76.12.6520

Cited by 7 publications

(1 citation statement)

References 16 publications

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“…The cells were harvested in the dark, cobalamins extracted with ethanol (80°C), separated by HPLC and radioactivity measured in fractions corresponding to marker cobalamins. Complementation studies were performed in ®broblasts fused with polyethyleneglycol (PEG) essentially as previously reported [9]; then methionine formation was measured as described above. Radioactively labelled compounds were purchased from Amersham, UK.…”

Section: Methodsmentioning

confidence: 99%

Methionine synthase deficiency without megaloblastic anaemia

Kvittingen

Spangen²,

Lindemans

et al. 1997

European Journal of Pediatrics

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Section: Methodsmentioning

confidence: 99%

Methionine synthase deficiency without megaloblastic anaemia

Kvittingen

Spangen²,

Lindemans

et al. 1997

European Journal of Pediatrics

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Studies on complementation in argininosuccinate synthetase and argininosuccinate lyase deficiencies in human fibroblasts

et al. 1981

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Complementation tests after polyethylene glycol fusion have been performed between 10 citrullinemic strains with argininosuccinate synthetase deficiency and between five strains with argininosuccinate lyase deficiency. No complementation was observed between the citrullinemic strains, while two groups of complementation were defined with the argininosuccinate lyase deficients strains. Since the restoration of activity was not total, we assumed that complementation is probably intragenic.

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Complementation analysis in Gaucher disease using single cell microassay techniques. Evidence for a single ?Gaucher gene?

Gravel

Leung

1983

Hum Genet

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Gaucher disease is a lysosomal storage disorder resulting from a deficiency of acid beta-glucosidase. Several clinical forms have been described, including infantile, juvenile, and adult onset variant. We have examined complementation in infantile and adult forms of Gaucher disease by monitoring enzyme activity in multinucleate cells produced by fusing skin fibroblasts from different patients in the presence of polyethylene glycol. beta-Glucosidase activity was monitored in lysates of individual multinucleate cells by a microassay method utilizing methylumbelliferyl-beta-D-glucoside as the substrate (normal: 1.3 +/- 0.12 x 10(-13) mol/h/cell). The microassay was linear with time up to 4 h, for up to 20 mononucleate cells, and for individual multinucleate cells containing up to 12 nuclei. Complementation was examined in 11 fibroblasts strains fused in all pairwise combinations. In no instance was there any clear indication of complementation (at least 10-15% of normal activity to adequately account for experimental variation) although there was an indication of marginal increases in some fusions. On the other hand, the expected 50% activity was obtained in "heterozygous" fusions (normal/mutant) for both types of clinical variants. Our results are consistent with a single gene, presumably the structural gene encoding the enzyme, responsible for at least the infantile and adult variants, and confirm the autosomal recessive nature of the disorder.

show abstract

Analysis of genetic complementation by whole-cell microtechniques in fibroblast heterokaryons.

Cited by 7 publications

References 16 publications

Methionine synthase deficiency without megaloblastic anaemia

Methionine synthase deficiency without megaloblastic anaemia

Studies on complementation in argininosuccinate synthetase and argininosuccinate lyase deficiencies in human fibroblasts

Complementation analysis in Gaucher disease using single cell microassay techniques. Evidence for a single ?Gaucher gene?

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