ISSN:1424-8581    

DOI:10.1159/000133330

出版商:S. Karger AG    

年代:1992

数据来源: Karger

ISSN:1424-8581    

DOI:10.1159/000133331

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

A method for the detection by electron microscopy of chromosome banding after in situ hybridization of small, nonradioactive DNA sequences is described. Typical high-resolution G-banding is produced by adding 5-bromodeoxyuridine (BrdU) during the last part of the S-phase and by applying a monoclonal antibody against the BrdU-substituted chromosome segments, followed by the addition of protein G, but no further treatment. A protocol for in situ hybridization of small, single-copy biotinylated DNA sequences and their detection by immunogold tagging on banded chromosomes is also described. This combined approach permits high-resolution mapping of small DNA sequences and should be useful in discriminating between neighboring DNA fragments.

ISSN:1424-8581    

DOI:10.1159/000133332

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Fluorescence in situ hybridization (FISH) was applied to detect the copy number of the retinoblastoma (RB1) tumor suppressor gene in metaphase chromosomes and interphase nuclei. We used 14 λ phage clones spanning the whole RB1 gene region as a probe and obtained a specific hybridization signal in normal metaphase chromosomes at 13q14. Normal interphase nuclei showed two RB1 signals in about 90% of cases, whereas two cell lines with cytogenetically defined deletions involving the RB1 gene showed only one hybridization signal in about 80% of the nuclei. Analogous changes were detected in metaphase chromosomes. Multicolor FISH with subsets of the phage clones allowed visualization of subregions within the 200-kb gene in interphase nuclei. Analysis of clinical breast cancer samples showed that most of the cells contained two copies of the RB1 gene, even when restriction fragment length polymorphism analysis showed loss of heterozygosity (LOH) at the RB1 locus. This indicates that LOH at the RB1 locus in breast cancer cells probably involves mechanisms other than physical deletion

ISSN:1424-8581    

DOI:10.1159/000133333

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

X-linked liver glycogenosis (XLG) is a glycogen storage disorder resulting from deficient activity of phosphorylase kinase (PHK). PHK consists of four different subunits: α, β, γ, and δ. Several genes encoding PHK subunits have been cloned and localized, but only the muscle α-subunit (PHKA) gene has been assigned to the X chromosome, in the region Xq12→q13. However, we have previously excluded the muscle PHKA gene as a candidate gene for the XLG mutation, as linkage analysis indicated that the mutation responsible for XLG is located in Xp22 and not in Xq12→q13. We report here the chromosomal localization by in situ hybridization of a liver PHKA gene to the distal region of chromosome Xp. Strong hybridization signals were observed on the distal part of the short arm of a chromosome identified as the X chromosome by cohybridization with an X chromosome-specific centromeric probe. The localization of this gene in the same chromosomal region as the disease gene responsible for XLG suggests that the liver PHKA gene is a highly likely candidate gene for the XLG

ISSN:1424-8581    

DOI:10.1159/000133334

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Through in situ hybridization of a cDNA probe to metaphase chromosomes, we localized the gene for the human urokinase receptor (PLAUR) on chromosome 19. RBG-banding permitted subchromosomal localization of the PLAUR gene to 19q13.

ISSN:1424-8581    

DOI:10.1159/000133335

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

By somatic cell fusion studies between noninvasive mouse T-lymphoma cells and invasive human activated normal T-cells we have previously shown that the genetic information responsible for the induction of invasive and metastatic potential in interspecies T-cell hybrids is located on human chromosome 7. Apparently, genes derived from normal activated T-cells are dominantly expressed in the hybrids and control the invasive and, as a consequence, metastatic potential of these T-lymphoma cells. To sublocalize the invasion-inducing locus on chromosome 7 we have generated hybrids that harbor only specific regions of human chromosome 7 with or without a small fragment of human chromosome 21. Analysis of these hybrids revealed that the invasion-inducing locus maps to 7p12→cen. The human DNA complement of the hybrids was confirmed by Southern blot analysis using a large panel of chromosome 7-specific DNA probes. Several of these genes could be further sublocalized. These included: ARAF2 to 7p12→cen, D7S21 to 7pter→p12, ACTB to 7p15→p12, EGFR to 7p12, MDH2 to 7cen→q22, and PDGFA to

ISSN:1424-8581    

DOI:10.1159/000133336

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

The human PDGFA gene has been mapped previously to two different sites on chromosome 7 (7p21→p22 and 7q11.2→q21.1). Using in situ hybridization and human × mouse somatic cell hybrid lines we present data which confirm the localization of PDGFA to the terminal short arm of chromosome 7, at

ISSN:1424-8581    

DOI:10.1159/000133337

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Aspartylglucosaminuria (AGU) is a recessive autosomally inherited lysosomal storage disorder due to deficiency of the enzyme aspartylglucosaminidase (AGA). The structural gene for this human enzyme (AGA) has been assigned to the region 4q21→qter. We determined the AGA activity in cultured fibroblasts of a girl with a 46, XX, del(4)(q33) karyotype. The results indicate that the girl is a hemizygote for AGA, permitting the assignment of human AGA to the region 4q33→q

ISSN:1424-8581    

DOI:10.1159/000133338

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

The chromosomal localization of the human gene coding for connexin 32 (GJB1) was determined by in situ suppression hybridization (ISSH). The results allowed assignment of the gene to band Xq13, thus refining previous localization data obtained by means of somatic cell hybrid analysis.

ISSN:1424-8581    

DOI:10.1159/000133339

出版商:S. Karger AG    

年代:1992

数据来源: Karger