Southern Analysis of the Lesch-Nyhan Locus in Man

SummaryProbands from 15 unrelated families with hereditary protein S deficiency type I, that is having a plasma total protein S concentration fifty percent of normal, were screened for abnormalities in their protein S genes by Southern analysis. Two probands were found to have a deviating DNA pattern with the restriction enzyme Mspl. In the two patients the alteration concerned the disappearance of a Mspl restriction site, CCGG, giving rise to an additional hybridizing Mspl fragment.Analysis of relatives of both probands showed that in one family the mutation does not co-segregate with the phenotype of reduced plasma protein S. In the family of the other proband, however, complete linkage between the mutated gene pattern and the reduced total protein S concentration was found: 12 heterozygous relatives showed the additional Mspl fragment but none of the investigated 26 normal members of the family. The mutation is shown to reside in the PSβ gene, the inactive protein S gene. The cause of type I protein S deficiency, a defect PSα gene has escaped detection by Southern analysis. No recombination has occurred between the PSα gene and the PSβ gene in 23 informative meioses. This suggests that the two protein S genes, located near the centromere of chromosome 3, are within 4 centiMorgan of each other.

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A southern analysis of Rh blood group genes: association between restriction fragment length polymorphism patterns and Rh serotypes

Blood ◽

10.1182/blood.v83.2.566.566 ◽

1994 ◽

Vol 83 (2) ◽

pp. 566-572 ◽

Cited By ~ 15

Author(s):

CA Hyland ◽

LC Wolter ◽

YW Liew ◽

A Saul

Keyword(s):

Restriction Fragment Length Polymorphism ◽

Blood Group ◽

Restriction Fragment ◽

Length Polymorphism ◽

Fragment Length ◽

Fragment Length Polymorphism ◽

Gene Dosage ◽

Southern Analysis ◽

Rh Blood Group ◽

Restriction Fragment Length

Abstract Polymorphisms within the Rh blood group system have been defined by serologic agglutination methods, but have not yet been defined at the DNA level. Two closely related genes associated with the Rh D antigen and with the Rh C/c and E/e antigens have been cloned. We used a Southern analysis incorporating probes to the 5′ and 3′ regions of the Rh C, E gene and D gene to identify polymorphisms associated with Rh C/c and E/e antigens, respectively. The D gene dosage could be determined by comparing the relative intensities of the D bands with bands from the 5′ and 3′ region of the Rh C, E gene. The concordance between restriction fragment length polymorphism (RFLP) patterns and serologic phenotypes for 102 randomly selected blood donors was 100% for C, e, and D, 94.8% for c, and 94.3% for E. The data are consistent with the sequences encoding the C/c epitopes residing on the 5′ side of those for the E/e epitopes. All samples discordant for the 3′ probe and E had the cE (r″) serotype. These data show that the gene coding for the cE serotype is different in Rh-positive and -negative individuals. The study demonstrates that Rh DNA typing, including D gene dosage measurements and Rh gene haplotyping, may supplement traditional serotyping methods in transfusion medicine.

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Islands of complex DNA are widespread in Drosophila centric heterochromatin.

Genetics ◽

10.1093/genetics/141.1.283 ◽

1995 ◽

Vol 141 (1) ◽

pp. 283-303

Author(s):

M H Le ◽

D Duricka ◽

G H Karpen

Keyword(s):

Dna Sequences ◽

Structure And Function ◽

Southern Analysis ◽

Cloning And Sequencing ◽

Pulsed Field ◽

Drosophila Heterochromatin ◽

And Function ◽

Heterochromatin Structure ◽

Derivatives Of ◽

Eukaryotic Genomes

Abstract Heterochromatin is a ubiquitous yet poorly understood component of multicellular eukaryotic genomes. Major gaps exist in our knowledge of the nature and overall organization of DNA sequences present in heterochromatin. We have investigated the molecular structure of the 1 Mb of centric heterochromatin in the Drosophila minichromosome Dp1187. A genetic screen of irradiated minichromosomes yielded rearranged derivatives of Dp1187 whose structures were determined by pulsed-field Southern analysis and PCR. Three Dp1187 deletion derivatives and an inversion had one breakpoint in the euchromatin and one in the heterochromatin, providing direct molecular access to previously inaccessible parts of the heterochromatin. End-probed pulsed-field restriction mapping revealed the presence of at least three "islands" of complex DNA, Tahiti, Moorea, and Bora Bora, constituting approximately one half of the Dp1187 heterochromatin. Pulsed-field Southern analysis demonstrated that Drosophila heterochromatin in general is composed of alternating blocks of complex DNA and simple satellite DNA. Cloning and sequencing of a small part of one island, Tahiti, demonstrated the presence of a retroposon. The implications of these findings to heterochromatin structure and function are discussed.

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Homology-Dependent Silencing of the SC3 Gene in Schizophyllum commune

Genetics ◽

10.1093/genetics/147.2.589 ◽

1997 ◽

Vol 147 (2) ◽

pp. 589-596 ◽

Cited By ~ 1

Author(s):

Theo A Schuurs ◽

Eveline A M Schaeffer ◽

Joseph G H Wessels

Keyword(s):

Gene Silencing ◽

Genomic Dna ◽

Type Strain ◽

Cytosine Methylation ◽

Wild Type Strain ◽

Schizophyllum Commune ◽

Southern Analysis ◽

Transcriptional Level ◽

Wild Type

After introduction of extra copies of the SC3 hydrophobin gene into a wild-type strain of Schizophyllum commune, gene silencing was observed acting on both endogenous and introduced SC3 genes in primary vegetative transformants. Nuclear run-on experiments indicated that silencing acted at the transcriptional level. Southern analysis revealed that cytosine methylation of genomic DNA occurred. Moreover, SC3 silencing was suppressed by exposure to 5-azacytidine during growth. After growth of SC3-suppressed colonies from homogenized mycelium or from colonies stored at 4°, SC3 transcription was restored. However, after prolonged growth SC3 silencing was again observed. Introduction of a promoterless SC3 fragment into wild type gave less SC3 silencing.

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Performance of an Operating High Energy Physics Data Grid: DØSAR-Grid

International Journal of Modern Physics A ◽

10.1142/s0217751x05027850 ◽

2005 ◽

Vol 20 (16) ◽

pp. 3874-3876 ◽

Cited By ~ 2

Author(s):

B. Abbott ◽

P. Baringer ◽

T. Bolton ◽

Z. Greenwood ◽

E. Gregores ◽

...

Keyword(s):

Geographical Distribution ◽

High Energy Physics ◽

High Energy ◽

End Users ◽

Southern Analysis ◽

Data Analyses ◽

Use Of Technology ◽

Computing Grid ◽

Physics Experiments ◽

Energy Physics

The DØ experiment at Fermilab's Tevatron will record several petabytes of data over the next five years in pursuing the goals of understanding nature and searching for the origin of mass. Computing resources required to analyze these data far exceed capabilities of any one institution. Moreover, the widely scattered geographical distribution of DØ collaborators poses further serious difficulties for optimal use of human and computing resources. These difficulties will exacerbate in future high energy physics experiments, like the LHC. The computing grid has long been recognized as a solution to these problems. This technology is being made a more immediate reality to end users in DØ by developing a grid in the DØ Southern Analysis Region (DØSAR), DØSAR-Grid, using all available resources within it and a home-grown local task manager, McFarm. We will present the architecture in which the DØSAR-Grid is implemented, the use of technology and the functionality of the grid, and the experience from operating the grid in simulation, reprocessing and data analyses for a currently running HEP experiment.

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Elimination of micronuclear specific DNA sequences early in anlagen development

Molecular and Cellular Biology ◽

10.1128/mcb.5.1.93-98.1985 ◽

1985 ◽

Vol 5 (1) ◽

pp. 93-98

Author(s):

C F Brunk ◽

R K Conover

Keyword(s):

Developmental Stage ◽

Dna Sequences ◽

Developmental Stages ◽

Tetrahymena Thermophila ◽

Southern Analysis ◽

Limited Sequence

After conjugation in Tetrahymena thermophila, the old macronuclei degenerate, and new macronuclei (anlagen) develop. During anlagen development a number of DNA sequences found in the micronuclear genome (micronuclear limited sequences) are eliminated from the anlagen. A cloned copy of a repetitive micronuclear limited sequence has been used to determine the developmental stage at which micronuclear limited sequences are eliminated. DNAs from anlagen of various developmental stages were examined by Southern analysis. It was found that micronuclear limited sequences are present in 4C anlagen and essentially absent in 8C and 16C anlagen. The precipitous loss of these sequences in the 8C anlagen rules out under-replication as the mechanism for the loss and suggests that these sequences are specifically degraded early during anlagen development.

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