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  • 1
    Electronic Resource
    Electronic Resource
    Long-range structure of H-ras 1-selected transgenomes (1989)
    Springer
    Somatic cell and molecular genetics 15 (1989), S. 229-235 
    Details
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have used chromosome-mediated gene transfer (CMGT) and whole cell fusion to derive human-mouse hybrid cells carrying reduced human chromosomes 11, by selecting for expression of the transforming H-ras 1 oncogene. To realize the full potential of these somatic cell genetic techniques as resources for enriched DNA probe isolation and the fine structure mapping of chromosomes, the nature of any molecular rearrangements that may accompany the process of DNA transfer must be understood. We have analyzed the long-range structure of our transgenomes by pulsed field gel electrophoresis (PFGE) and show here that, whereas during cell fusion several megabase pairs (Mb) of DNA can be transferred intact, multiple rearrangements of DNA accompany CMGT even in transgenomes where other methods of analysis gave no indication of such molecular scrambling.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01534873
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  • 2
    Electronic Resource
    Electronic Resource
    Rapid and quantitative detection of unique sequence donor DNA in extracts of cultured mammalian cells: An aid to chromosome mapping (1985)
    Springer
    Somatic cell and molecular genetics 11 (1985), S. 445-454 
    Details
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A rapid and highly sensitive method for screening the human DNA content of hybrid or transfected mammalian cells is described. Transfectants containing as little as 200 kb of otherwise undefined human DNA can be readily detected in a background of mouse chromatin. At the highest stringency, single-copy sequences can be detected. Large numbers of independent gene-transfer products are easily screened, making the method ideally suited to the identification of rare, but otherwise unselectable, events. The method does not rely upon the expression of the gene sequence of interest; the sole proviso is the availability of an appropriate DNA probe for the chromosomal region or locus of interest.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01534838
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  • 3
    Electronic Resource
    Electronic Resource
    Analysis of WAGR deletions and related translocations with gene-specific DNA probes, using FACS-selected cell hybrids (1988)
    Springer
    Somatic cell and molecular genetics 14 (1988), S. 21-30 
    Details
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We used the fluorescence-activated cell sorter (FACS) to select a series of somatic cell hybrids with deleted or translocated chromosome 11 segregated from its normal homolog. Analysis of these cell hybrids with gene-specific probes and for cell-surface marker expression has allowed us to order the markers and define a smallest region of overlap (SRO) for deletions associated with the WAGR (Wilms' tumor, aniridia, genitourinary abnormalities, and mental retardation) region of chromosome 11. Two translocation breakpoints in 11 p13 (one associated with familial aniridia and one with a sporadic case of congenital renal dysfunction resulting from urethral and ureteral atresia) map within this SRO.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01535046
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  • 4
    Electronic Resource
    Electronic Resource
    SV40-mediated tumor selection and chromosome transfer to enrich for cystic fibrosis region (1990)
    Springer
    Somatic cell and molecular genetics 16 (1990), S. 29-38 
    Details
    ISSN: 1572-9931
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The somatic cell hybrid C121, with chromosome 7 as its sole human component, arose when mouse macrophages were immortalized by fusion with SV40-transformed human fibroblasts. The transforming SV40 genomes are integrated at 7q31-7q35. We show that hybrids with a reduced chromosome 7 component, but which retain markers linked to the cystic fibrosis locus, can be generated by direct in vivo tumor selection or following chromosome-mediated gene transfer and SV40-mediated cellular transformation. Our methods for chromosome fragmentation and fine-structure mapping can now be applied to the substantial number of SV40-transformed human cell lines, with independent chromosomal integration sites, already available. Our results also suggest that expression of human epidermal growth factor receptor augments the tumorigenic potential of the SV40-transformed C121 hybrid.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01650477
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  • 5
    Electronic Resource
    Electronic Resource
    EagI and NotI linking clones from human chromosomes 11 and Xp (1996)
    Springer
    Human genetics 〈Berlin〉 97 (1996), S. 742-749 
    Details
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract EagI and NotI linking libraries were prepared in the lambda vector, EMBL5, from the mouse-human somatic cell hybrid 1W1LA4.9, which contains human chromosomes 11 and Xp as the only human component. Individual clones containing human DNA were isolated by their ability to hybridise with total human DNA and digested with SalI and EcoRI to identify the human insert size and single-copy fragments. The mean (± SD) insert sizes of the EagI and NotI clones were 18.3 ± 3.2 kb and 16.6 ± 3.6 kb, respectively. Regional localisation of 66 clones (52 EagI, 14 NotI) was achieved using a panel of 20 somatic cell hybrids that contained different overlapping deletions of chromosomes 11 or Xp. Thirty-nine clones (36 EagI, 3 NotI) were localised to chromosome 11; 17 of these were clustered in 11q13 and another nine were clustered in 11q14–q23.1. Twenty-seven clones (16 EagI, 11 NotI) were localised to Xp and 10 of these were clustered in Xp11. The 66 clones were assessed for seven different microsatellite repetitive sequences; restriction fragment length polymorphisms for five clones from 11q13 were also identified. These EagI and NotI clones, which supplement those previously mapped to chromosome 11 and Xp, should facilitate the generation of more detailed maps and the identification of genes that are associated with CpG-rich islands.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004390050130
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  • 6
    Electronic Resource
    Electronic Resource
    EagI andNotI linking clones from human chromosomes 11 and Xp (1996)
    Springer
    Human genetics 〈Berlin〉 97 (1996), S. 742-749 
    Details
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract EagI andNotI linking libraries were prepared in the lambda vector, EMBL5, from the mouse-human somatic cell hybrid 1W1LA4.9, which contains human chromosomes 11 and Xp as the only human component. Individual clones containing human DNA were isolated by their ability to hybridise with total human DNA and digested withSalI andEcoRI to identify the human insert size and single-copy fragments. The mean (± SD) insert sizes of theEagI andNotI clones were 18.3 ± 3.2 kb and 16.6 ± 3.6 kb, respectively. Regional localisation of 66 clones (52EagI, 14NotI) was achieved using a panel of 20 somatic cell hybrids that contained different overlapping deletions of chromosomes 11 or Xp. Thirty-nine clones (36EagI, 3NotI) were localised to chromosome 11; 17 of these were clustered in llg13 and another nine were clustered in 11ql4–q23.1. Twenty-seven clones (16EagI, 11NotI) were localised to Xp and 10 of these were clustered in Xpll. The 66 clones were assessed for seven different microsatellite repetitive sequences; restriction fragment length polymorphisms for five clones from 11q13 were also identified. TheseEagI andNotI clones, which supplement those previously mapped to chromosome 11 and Xp, should facilitate the generation of more detailed maps and the identification of genes that are associated with CpG-rich islands.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02346183
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  • 7
    Electronic Resource
    Electronic Resource
    Rapid isolation of moderate and highly polymorphic DNA fragments mapping close to WT (Wilms' tumour) and AN2 (aniridia) on chromosome 11 (1989)
    Springer
    Human genetics 〈Berlin〉 81 (1989), S. 349-352 
    Details
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Genes implicated in the development of Wilms' tumour (WT) and aniridia (AN2) have been localised to a subregion of band p13 on chromosome 11 by molecular and cytogenetic characterisation of WAGR syndrome patients carrying variable constitutional deletions. Polymorphic markers for the region would be valuable for linkage analysis in the familial forms of both Wilms' tumour and aniridia, as well as for studying somatic rearrangements of chromosome 11 in a variety of tumour types. Here we describe the isolation and characterisation of three frequently polymorphic arbitrary DNA fragments that map proximal to the AN2 and WT loci.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00283689
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  • 8
    Electronic Resource
    Electronic Resource
    Isolation and characterization of the mouse translin-associated protein X (Trax) gene (2000)
    Springer
    Mammalian genome 11 (2000), S. 395-398 
    Details
    ISSN: 1432-1777
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s003350010074
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  • 9
    Electronic Resource
    Electronic Resource
    Distribution of Alu and L1 repeats in human YAC recombinants (1992)
    Springer
    Mammalian genome 3 (1992), S. 661-668 
    Details
    ISSN: 1432-1777
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Evidence is accumulating that the two major families of interspersed repeated human DNA sequences, Alu and L1, are not randomly distributed. However, only limited information is available on their relative long-range distribution. We have analyzed a set of randomly selected, human Chromosome (Chr) 11-specific YAC recombinants constituting a total length of about 2 Mbp for the local and global distribution of Alu and L1 repeats: the data show a strong asymmetry in the distribution of these two repeat classes and give weight, at the long-range molecular level, to previous studies indicating their partition in the human genome; they also suggest a strong tendency for L1 repeats to cluster, with a higher proportion of full-length elements than expected.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00444360
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  • 10
    Electronic Resource
    Electronic Resource
    Enhancing the efficiency of introducing precise mutations into the mouse genome by hit and run gene targeting (2000)
    Springer
    Transgenic research 9 (2000), S. 55-66 
    Details
    ISSN: 1573-9368
    Keywords: cystic fibrosis ; ES cells ; gene inactivation ; gene targeting ; hit and run
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The creation of precise clinical mutations by gene targeting is important in elucidating disease pathogenesis using mouse models. 'Hit and run' gene targeting is an elegant method to achieve this goal. This uses first a positive selection to introduce the targeting vector carrying the required mutation and then a negative selection to identify clones which have removed vector and wild-type sequences by intrachromosomal recombination. However, this approach has only been successfully used in a handful of cases. We used this procedure to introduce precise clinical mutations into the exon 10 region of the cystic fibrosis transmembrane conductance regulator (Cftr) gene. Using a CMV promoter driven hygromycin/thymidine kinase (hyg/tk) fusion gene as both our dominant and negative selectable marker, we targeted the Cftr locus very efficiently but only identified false runs after the negative selection step. This defect in thymidine kinase induced toxicity to gancyclovir correlated with methylation of the transgene. Consequently we devised a stringent screening procedure to select only true 'run' clones. Unfortunately these 'run' clones had lost the mutation so we altered the vector design to bias the run step to retain the mutation and used a different tk selection cassette with a HSVtk promoter sequence. This new vector design allowed both efficient 'hit and run' for two cystic fibrosis (CF) mutations with no false positives and successful germline transmission of the novel G480C missense mutation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008915026660
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