摘要:

We provide fast, simple, one-step procedures for sequence-specific detection of nucleic acids in situ. Tandem repeat sequences in DNA are stained within 30 min, and mRNA is stained within 2 h. The procedures are based on the incorporation of the newly available fluorescein-labeled dUTP into DNA synthesized in situ by primed in situ labeling, with denatured fragments of cloned DNA or oligonucleotides as primers. The extreme speed and simplicity of the reaction make it attractive for automatization in routine laboratory procedures and opens up new diagnostic possibilities.

ISSN:1424-8581    

DOI:10.1159/000133281

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

A double-labeling approach was applied to nonisotopic in situ hybridization with individual cosmid and plasmid clones, using digoxigenin or biotin as label and a combination of two separate enzymatic labeling methods. Probe labeling was achieved by nick translation, followed by tailing of the probe by terminal deoxynucleotidyl transferase. The double-labeling method, in conjunction with an improved detection protocol, provides for a higher signal intensity than that obtainable with single-labeled probes.

ISSN:1424-8581    

DOI:10.1159/000133282

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Marker chromosomes contain potentially valuable information about breakpoints in cancer. However, routine banding procedures, by themselves, provide only limited information about the identity of marker chromosomes. In this study, the use of fluorescence in situ hybridization (FISH) with chromosome-specific centromeric probes and whole-chromosome-specific DNA libraries greatly enhanced the identification of 10 marker chromosomes in the primary prostatic cancer cell line PPC-1. Centromeric probes for chromosomes 1, 2, 3, 4, 10, 12, and 17 and whole-chromosome paint libraries for chromosomes 1, 2, 3, 4, 8, and 12, in conjunction with analysis of G-banded metaphases, allowed the major portion(s) of these 10 PPC-1 marker chromosomes to be defined. The results increase the number of identifiable chromosomal breakpoints in this cell line from 9 to 28 sites.

ISSN:1424-8581    

DOI:10.1159/000133283

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

A patient with the CREST syndrome of scleroderma was found to carry a mosaicism for a supernumerary microchromosome. The microchromosome was ∼1 µm in size and present in over half of the lymphocyte metaphases examined. It bound centromeric proteins specifically recognized by CREST autoimmune sera (including the patient’s serum). In situ hybridization with a panel of chromosome-specific α-satellite probes showed that the microchromosome was derived from chromosome 11 most or all of its chromatin consisting of the chromosome 11 subset of α-satellite DNA. It had no detectable telomeric sequences. Microchromosomes observed by electron microscopy had no visible free ends. The chromatin looked exactly the same as it did in normal chromosomes. Although we have no direct evidence for a circular structure, we conclude that the microchromosome originated by an interstitial deletion including the α-satellite DNA sequences and subsequent ring formation. The newly formed chromosomal element proved to be relatively stable somatically and was transmitted through meiosis. Since it possesses at least some structural and functional features of a centromeric region, the microchromosome can be thought of as an isolated cen

ISSN:1424-8581    

DOI:10.1159/000133284

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Sperm chromosome complements were analysed in two men who were heterozygous carriers of reciprocal translocations. A total of 363 sperm were karyotyped after in vitro penetration of hamster oocytes, including 180 sperm from a male with a t(1;9)(q22;q31) and 183 from a male with a t(16;19)(q11.1;q13.3). All possible 2:2 and 3:1 meiotic segregations were observed for both translocations. The frequencies of alternate, adjacent 1 adjacent 2, and 3:1 segregations were 46 %, 38 %, 13 %, and 4 % for the t(1;9) and 40%, 28 %, 31 %, and 1 % for the t(16;19), respectively. Within the alternate segregation group, the number of normal sperm was not significantly different from the number of sperm carrying a balanced form of the translocation for either of the translocations, as expected. There was no evidence for an interchromosomal effect of either translocation, since the frequencies of numerical abnormalities unrelated to the translocation were within the normal range observed in sperm from control donors. The percentage of sperm with an unbalanced form of the translocation was 54% for the t(1;9) and 61 % for the t(16;19).

ISSN:1424-8581    

DOI:10.1159/000133285

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

A cDNA for a new catalytic subunit (Cγ) of the cAMP-dependent protein kinase (PKA) was recently isolated from a human testis cDNA library. This subunit was shown to be expressed only in testis, and has so far not been demonstrated in other species. In the present study, we have determined the chromosomal localization of this gene employing a cDNA for Cγ as a probe. Southern blot analysis of genomic DNA from human × mouse somatic cell hybrids allowed us to assign this gene (PRKACG) to human chromosome 9. In situ hybridization to metaphase chromosomes confirmed the somatic cell hybrid data and regionally mapped the Cγ gene of PKA to human chromosome 9

ISSN:1424-8581    

DOI:10.1159/000133286

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

We have constructed a linkage map of eight RFLP markers located on chromosome 11 q in the region of the dopamine D2 receptor gene (DRD2) recognized by probe hD2Gl. Abnormalities in dopaminergic neurotransmission mediated by this receptor have been implicated in several psychiatric disorders. The map was generated using six large reference families (from 294 to 419 individuals per locus), which are largely independent of the CEPH families, primarily using the LINKMAP and ILINK programs of the LINKAGE package of Lathrop and Lalouel. The most likely order and recombination frequencies are: cen-DllS146-INT2-D11S36-(DllS84, STMY)-DRD2-D11S29-PBGD-qter 0.001 0.213 0.123 0.001 0.158 0.035 0.093 The relative order of Dl 1S84-STMY, DRD2-D11S29, and D11S146-INT2 could not be resolved reliably. There were no significant sex differences in recombination frequency. We introduce here a version of LINKMAP adapted to run under distributed parallel processing (LINDA-LINKMAP). Using pairwise analyses, we have also placed D11S421 proximal to this group.

ISSN:1424-8581    

DOI:10.1159/000133287

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

The human muscle adenine nucleotide translocator gene (ANT1) was previously assigned to chromosome 4. The gene has now been further localized to the long arm of chromosome 4 at 4q35 by fluorescence in situ hybridization. This result confirms the previous assignment and precisely maps the gene to a specific chromosome band.

ISSN:1424-8581    

DOI:10.1159/000133288

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

Human apolipoprotein H (APOH) is associated with lipoprotein present in plasma. It has been shown that APOH has structural similarities with the regulation of complement activation (RCA) protein superfamily and is involved in phospholipid binding interactions on platelets and as an autoantigen in complex with anionic phospholipids. Nevertheless, additional functional studies are necessary to establish the physiological role of APOH. By hybridizing a cDNA probe for APOH to a panel of somatic cell hybrids, we show that the structural locus for this protein maps to 17q23→qter and is therefore not part of the RCA cluster on chromosome 1. The site of biosynthesis for APOH was established by Northern blot analysis. Hybridization of the APOH cDNA probe to total liver RNA identified a transcript of approximately 1.5 k

ISSN:1424-8581    

DOI:10.1159/000133289

出版商:S. Karger AG    

年代:1992

数据来源: Karger

摘要:

The gene for the fibroblast growth factor receptor BEK was assigned to human chromosome 10 by applying polymerase chain reaction techniques to DNAs from a panel of human × rodent somatic cell hybrids. The gene was further localized to 10q25.3→q26 by in situ hybridizati

ISSN:1424-8581    

DOI:10.1159/000133290

出版商:S. Karger AG    

年代:1992

数据来源: Karger