High-Resolution Chromosome Sorting and DNA Spot-Blot Analysis Assign McArdle's Syndrome to Chromosome 11

Lebo, Roger V.; Gorin, Fredric A; Fletterick, Robert J.; Kao, Fa Ten; Cheung, Michael K.; Bruce, Barry D.; Kan, Yuet Wai

doi:10.1126/science.6587566

Cited by 222 publications

(91 citation statements)

References 14 publications

(16 reference statements)

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“…2). This separation pattern essentially coincides with a projection of the dual parameter flow histogram to the H33258-axis reported by Lebo et al (13). The chromosome types contained in each fraction were determined by spot-blot hybridization using chromosome-specific DNA markers as probes.…”

Section: High Resolution Flow Analysis and Sorting Of Human Chromosomessupporting

confidence: 85%

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Isolation of giant DNA fragments from flow‐sorted human chromosomes

et al. 1990

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We have established a method using a conventional cell sorter equipped with a single argon laser to sort intact human chromosomes that can be used as a source for the production of giant DNA fragments. Various improvements were made to both the equipment and sorting method to enhance the sorting resolution and avoid destruction of chromosomal DNA. Using this improved method chromosomes 21 and 22 were sorted from the B-lymphoblastoid line GM00130B, digested with the rare cutting restriction endonuclease NotI, and analyzed by pulsed field gel electrophoresis followed by Southern hybridization using the AZu repetitive sequence as a probe. More than 25 discrete NotI giant DNA fragments ranging from 50 kb to longer than 2.5 Mb were separated and the size distribution pattern was unique for each chromosome, indicating successful sorting of intact chromosomes. The cumulative size of these AZu-positive NotI DNA fragments were 22.7 Mb and 25.5 Mb for chromosomes 21 and 22, respectively. These values are 47% and 49% of the estimated size of chromosomes 21 (48 Mb) and 22 (52 Mb).

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Section: High Resolution Flow Analysis and Sorting Of Human Chromosomessupporting

confidence: 85%

“…By appropriately using either dye, 17 types of human chromosomes (1-8, 13-16, 18, 20-22, and X) can be sorted as single components. Chromosomes 3, 8,13,18,20,21, and 22 are essentially pure.…”

Section: High Resolution Flow Analysis and Sorting Of Human Chromosomesmentioning

confidence: 99%

Isolation of giant DNA fragments from flow‐sorted human chromosomes

et al. 1990

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“…Human chromosomes were isolated and sorted directly on nitrocellulose discs as previously described [14,15] and hybridized to labelled Gp IX cDNA.…”

Section: Chromosomal Localizationmentioning

confidence: 99%

Human platelet glycoprotein IX

1990

FEBS Letters

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Overlapping cDNAs encoding human platelet glycoprotein (Gp)IX were cloned from a human erythroleukemia cell λ gtll library. The possibly ‘full‐length’ cDNA of 896 base pairs (bp) includes an open reading frame (528 bp), both 5' (222 bp) and 3' (146 bp) noncoding regions, and a poly(A) tail. Translation predicts a signal peptide of 16 amino acids and a mature protein of 160 amino acids that includes a 24 amino acid leucinerich glycoprotein (LRG) segment. Southern blot analysis suggests the presence of a single copy of the Gp IX gene, and hybridization of Gp IX cDNA to sorted human chromosomes localizes the Gp IX gene to chromosome 3.

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“…For swine, the MHC region was found on chromosome 7 by in situ hybridization by Geffrotin et al (7). Flow sorting associated with spot blot hybridization is a powerful technique that recently allowed regional mapping of different human genes (4,(13)(14)(15). The efficiency of the method prompted us to determine if it was applicable to porcine gene localisation.…”

mentioning

confidence: 99%

Swine chromosomes: Flow sorting and spot blot hybridization

et al. 1986

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Flow cytometry analysis was applied to swine chromosomes prepared from phytohemagglutinin (PHA) stimulated peripheral blood lymphocytes. Flow karyotypes from both sexes and from t(3;7) translocation carrier females were obtained. A certain number of chromosome pairs could be assigned to various peaks. In fact, 13 peaks were observed for 18 autosomal pairs plus X and Y. Moreover, abnormalties owing to the t(3;7) translocation were readily observable.The number of base pairs for chromosomes associated with the various peaks was estimated by comparison with human flow karyotypes.The following four peaks were thus sorted: (1) the peak assumed to represent the translocated chromosome 7 plus the normals associated with it; (2) the corresponding peak from a normal swine; (3) the peak assumed to contain among others the normal chromosome 7; and finally (4) the peak corresponding to swine chromosome 1.Chromosomes of each peak were collected on Pall Biodyne membrane. Following appropriate denaturation and prehybridization, the four samples were hybridized with a human leucocyte antigen (HLA) class I32P‐labelled cDNA probe, representing most of the coding sequence of the HLA B7 gene. The results confirmed previous data from other techniques that assigned the swine MHC(SLA) to chromosome 7.Subsequently, sorted samples were hybridized with a porcine genomic Interferon alpha probe in order to confirm the mapping of this gene family on porcine chromosome 1.

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High-Resolution Chromosome Sorting and DNA Spot-Blot Analysis Assign McArdle's Syndrome to Chromosome 11

Cited by 222 publications

References 14 publications

Isolation of giant DNA fragments from flow‐sorted human chromosomes

Isolation of giant DNA fragments from flow‐sorted human chromosomes

Human platelet glycoprotein IX

Swine chromosomes: Flow sorting and spot blot hybridization

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