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J. Kumlien T. Labella G. Zehetner R. Vatcheva D. Nizetic H. Lehrach 《Mammalian genome》1994,5(6):365-371
The use of integrated mapping strategies involving bacterial, yeast, and rodent cells as hosts simplifies the construction of maps, which combine long-range order, high resolution, and easy access to the cloned DNA. Radiation-fusion hybrids offer a specially powerful long-range mapping system for human chromosomes. We describe here techniques for establishing a radiation-fusion hybrid map of Chromsome (Chr) 21 q and its integration with local information on YAC and cosmid positions. 相似文献
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F Muscatelli A P Monaco P N Goodfellow M C Hors-Cayla H Lehrach M Fontes 《Cytogenetics and cell genetics》1992,61(2):109-113
In order to isolate new probes from the juxtacentromeric region of the long arm of the human X chromosome, we used Alu-mediated polymerase chain reaction (Alu-PCR) products as probes to directly screen a chromosome X-specific gridded cosmid library. These Alu-PCR products were synthesized from radiation hybrids containing the loci DXS159, PGK1, and PGK1P1. This approach allowed us to select 18 cosmids capable of hybridizing with at least two Alu-PCR products. Four cosmids hybridized to more than three Alu-PCR products. Three of these four cosmids were contiguous, and the fourth was independent. Two cosmids that hybridized with two Alu-PCR products were further characterized. Physical mapping indicated that all of these clones are located in the expected region on Xq, confirming the validity of our approach. 相似文献
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Roger D. Cox Alison Hugill Alexandra Shedlovsky Janice K. Noveroske Steve Best Monica J. Justice Hans Lehrach William F. Dove 《Genomics》1999,57(3):333
Multiple alleles of the quaking (qk) gene have a variety of phenotypes ranging in severity from early embryonic death to viable dysmyelination. A previous study identified a candidate gene, QKI, that contains an RNA-binding domain and encodes at least three protein isoforms (QKI-5, -6 and -7). We have determined the genomic structure of QKI, identifying an additional alternative end in cDNAs. Further we have examined the exons and splice sites for mutations in the lethal allelesqkl-1, qkkt1, qkk2,andqkkt3.The mutation inqkl-1creates a splice site in the terminal exon of the QKI-6 isoform. Missense mutations in the KH domain and the QUA1 domains inqkk2andqkkt3,respectively, indicate that these domains are of critical functional importance. Although homozygotes for each ENU induced allele die as embryos, their phenotypes as viable compound heterozygotes with qkvdiffer. Compound heterozygousqkvanimals carryingqkkt1, qkk2,andqkkt3all exhibit a permanent quaking phenotype similar to that ofqkv/qkvanimals, whereasqkv/qkl-1animals exhibit only a transient quaking phenotype. Theqkl-1mutation eliminates the QKI-5 isoform, showing that this isoform plays a crucial role in embryonic survival. The transient quaking phenotype observed inqkv/qkl-1mice indicates that the QKI-6 and QKI-7 isoforms function primarily during myelination, but that QKI-5 may have a concentration-dependent role in early myelination. This mutational analysis demonstrates the power of series of alleles to examine the function of complex loci and suggests that additional mutant alleles of quaking could reveal additional functions of this complex gene. 相似文献