摘要:

Fluorescent in situ hybridization was used to assign 37 yeast artificial chromosomes (YACs) from the CEPH (Centre d’Etude du Polymorphisme Humain) libraries to R-banded metaphases stained with propidium iodide. The use of the whole yeast DNA as probe, allows rapid mapping and characterization of YAC clones. Of 37 YAC clones, 11 were found cytogenetically chimeric (30%). The YAC clones reported in this study can be used to isolate and characterize targeted regions involved in genetic diseases and cancer breakpoint

ISSN:1424-8581    

DOI:10.1159/000133611

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

We have developed a method that allows reliable construction of high quality FISH probes from yeast artificial chromosomes (YACs) based on the separation of YACs by pulsed-field gel electrophoresis and a rapid sequence-independent amplification procedure (SIA). These probes can be used to localize YACs on metaphase chromosomes and also with high efficiency, in interphase nuclei.

ISSN:1424-8581    

DOI:10.1159/000133612

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

Oligonucleotide primers specific for exons 4 and 5 sequences were used to amplify a unique 199-bp fragment in the human GSTM3 gene. Using DNA from a panel of somatic cell hybrids we assigned the GSTM3 locus to chromosome 1p.

ISSN:1424-8581    

DOI:10.1159/000133613

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

We have determined the chromosomal localizations of newly isolated cosmids by fluorescent in situ hybridization (FISH) on prometaphase R-banded chromosomes and have constructed a high-resolution cytogenetic map for human chromosome 8 with 416 cosmid markers, including 328 new markers and 88 reported previously. Of the 416 markers, 229 were mapped to the long arm of chromosome 8, 181 to the short arm, and 6 to the centromere. Although the clones were scattered throughout the chromosome, they were concentrated in R-positive bands. Since the estimated physical length of chromosome 8 is 135 Mb, the overall average distance between loci is 320 kb, but the average separation of loci on R-positive bands is nearly 130–200 kb. This cytogenetic map will serve as a resource for efforts to characterize chromosomal and molecular aberrations involved in cancers, to clone genes associated with hereditary diseases, and to construct a detailed physical map of large electrophoretic fragments and/or contiguous cosmids and yeast artificial chromosome

ISSN:1424-8581    

DOI:10.1159/000133614

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

Chromosome assignments of the genes for rat ceruloplasmin (CP) and metallothionein (MT) were performed by analyzing somatic cell hybrid DNAs with the polymerase chain reaction (PCR), using primers specific for rat genes. The genes for CP and MT were assigned to rat chromosomes 2 and 19, respectively.

ISSN:1424-8581    

DOI:10.1159/000133615

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

Replication banding patterns were induced in the chromosomes of Gastrotheca riobambae by treating cell cultures with 5-bromodeoxyuridine (BrdU) and deoxythymidine (dT). In particular, the time sequence of replication of the highly differentiated XY/XX sex chromosomes was meticulously analyzed throughout the S phase. In the homogametic female, BrdU/dT labeling revealed no evidence of asynchronous replication between euchromatic regions in the XX pair. Minor replication asynchronies between the two X chromosomes were restricted to the heterochromatic C-bands. These results are further proof that dosage compensation in Amphibia for X- or Z-linked genes does not occur by inactivation of one of the two (XX or ZZ) sex chromosomes in the cells of homogametic individuals. In the very large and almost completely heterochromatic Y chromosome of G. riobambae, the various C-bands comprise at least three different structural categories, resulting in a complex pattern of early- and late-replicating bands along the Y chromosome.

ISSN:1424-8581    

DOI:10.1159/000133616

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

The retinoblastoma susceptibility gene, which encodes a nuclear phosphoprotein (Rb), plays an important role in cell-cycle progression in both normal development and tumorigenesis. Using a Syrian hamster cDNA probe homologous to the human retinoblastoma gene (RB), we have localized the gene to chromosome band 15q13 by conventional in situ hybridization.

ISSN:1424-8581    

DOI:10.1159/000133617

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

To improve resolution for physical ordering of adjacent DNA loci, prophase chromosomes were used for multi-color fluorescent in situ hybridization (FISH). The prophase chromosomes were prepared from cultured lymphocytes by a thymidine synchronization, bromodeoxyuridine release technique and then treating the synchronized cultures with topoisomerase II inhibitors ICRF154 or ICRF193. Almost all mitotic figures exhibited highly elongated prophase chromosomes without significant reduction of the mitotic index. Using multi-color FISH with these prophase chromosomes, we were able to distinguish signals for loci separated by as little as 50 kb, and determine their orientation. Furthermore, using this prophase ordering system, we confirmed the linear order and defined the orientation of seven cosmid markers within a 360-kb region surrounding D1OS102, a locus that is closely linked to the disease locus in families segregating an allele causing multiple endocrine neoplasia IIA (MEN2A). This prophase FISH system, by rapidly and precisely providing the linear order of loci that are very close, can expedite construction of fine cytogenetic maps and contribute to positional-cloning studies in which the precise ordering of DNA loci in a target region is critical.

ISSN:1424-8581    

DOI:10.1159/000133618

出版商:S. Karger AG    

年代:1994

数据来源: Karger

ISSN:1424-8581    

DOI:10.1159/000133619

出版商:S. Karger AG    

年代:1994

数据来源: Karger

摘要:

Fifty-six chromosome 18-specifιc yeast artificial chromosomes (YAC) were isolated from a human monochromosomal somatic cell hybrid DNA library. Fluorescence in situ hybridization revealed that the clones were evenly distributed throughout the chromosome. The clones, with an average insert size of 250 kb, cover about 18 % of the euchromatic part of chromosome 18. Of 90 STS markers tested, 17 were represented in this YAC collection. Together with our previously reported set of unique chromosome 18-specific YACs, we now have available 111 regionally mapped, essentially nonchimeric, clones that provide more than 30% coverage and form a framework for the complete physical map

ISSN:1424-8581    

DOI:10.1159/000133620

出版商:S. Karger AG    

年代:1994

数据来源: Karger