PubMed · 7184272
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W Schnedl. 1982. [Methods in human genetics].. https://pubmed.ncbi.nlm.nih.gov/7184272/
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OBJECTIVE: To develop a simple method for detecting DNA probes directly on R-banded chromosomes. METHODS: After sixty-seven hours culture, human peripheral blood lymphocytes were synchronized for 5 to 6 hours by adding Hoechst 33258 and BUdR, and then arrested by standard cytogenetic procedures. The slides were mounted with 2 x SSC and exposed with a 20W UV light which was about 10cm above the slides for 20 min at 75 degrees C. The biotinylated probes, such as the cosmid and YAC clones on 5p specific region and pBamX7, were hybridized on to the slides. After washing, the slides were treated with avidin-FITC and amplified with additional layer of biotinylated anti-avidin and avidin-FITC, and counterstained with propidium iodide in an antifade solution. Fluorescent signals and R-bands were observed simultaneously under Olympus BX 60 fluorescence microscope equipped with a WIB filter. RESULTS: The chromosomal location of the greenish-yellow signals could be directly identified on the R-banded chromosome background. CONCLUSION: This method can serve as a rapid and precise system for chromosomal localization of DNA markers.
SPOCK, previously identified as testican, is a modular proteoglycan that carries both chondroitin and heparan sulfate glycosaminoglycan side chains. The overall genomic organization has been established. The SPOCK gene spans at least 70 kb and is composed of 11 exons: the first half of the gene is dramatically expanded, but the second half is more compact. In situ hybridization and YAC mapping independently linked the SPOCK gene to 5q31, a region containing an impressive number of genes encoding growth factors, cytokines, and neurotransmitter and hormone receptors. The gene is located between the IL9 and the EGR1 genes, bordering the smallest commonly deleted region of chromosome 5.
Cytogenetic and molecular analyses were performed on three cellular (atypical) congenital mesoblastic nephromas (CMNs). Two cases had trisomy 11; in one, it was the sole karyotypic abnormality, and the other had additional numerical changes as well as an isochromosome for the long arm of chromosome 1. Markers for the 11p13 and 11p15 loci were present in three copies in these two CMNs. In the third CMN, two apparently normal copies of chromosome 11 were present together with additional numerical and structural chromosome changes. Because loss of heterozygosity was observed for both 11p13 and 11p15 markers, we assume that mitotic recombination occurred. Duplication and loss of imprinting of genes at 11p15 has also been observed frequently in Wilms' tumor. We therefore propose that CMN and Wilms' tumor might share common genetic pathways.