RESUMEN
We have used X-ray irradiation and cell fusion to generate somatic cell hybrids containing fragments of human chromosome 10. Our experiments were directed towards isolating the region of the MEN2A gene in hybrids and to use those as the source of DNA for cloning and mapping new markers from near the MEN2A locus. A number of hybrid clones containing human sequences that are tightly linked to the MEN2A gene were identified. Some 25% of our hybrids, however, proved to contain more than one human chromosome 10-derived fragment or showed evidence of deletions and/or rearrangements. A detailed analysis of the human content of X-ray irradiation hybrids is required to assess the integrity and number of human fragments retained. Despite retention of multiple human-derived fragments, these hybrids will prove useful as cloning and mapping resources.
Asunto(s)
Cromosomas Humanos Par 10/ultraestructura , ADN/ultraestructura , Células Híbridas/ultraestructura , Neoplasia Endocrina Múltiple/ultraestructura , Animales , Fusión Celular/efectos de la radiación , Línea Celular , Mapeo Cromosómico , Cromosomas Humanos Par 10/análisis , Cromosomas Humanos Par 10/efectos de la radiación , Cricetinae , Cricetulus , ADN/genética , ADN/efectos de la radiación , Técnica del Anticuerpo Fluorescente , Marcadores Genéticos/análisis , Humanos , Células Híbridas/análisis , Células Híbridas/efectos de la radiación , Neoplasia Endocrina Múltiple/análisis , Neoplasia Endocrina Múltiple/genética , Hibridación de Ácido Nucleico , Polimorfismo de Longitud del Fragmento de Restricción , Rayos X , Cromosoma Y/análisis , Cromosoma Y/efectos de la radiación , Cromosoma Y/ultraestructuraRESUMEN
There are at least three transcriptionally active human ADP/ATP translocase genes. We have isolated seven ADP/ATP translocase pseudogenes from recombinant human genomic libraries. Each pseudogene sequence had more than 85% identity with the sequence of the human ADP/ATP translocase cDNA derived from fibroblast mRNA, but each had mutations that precluded synthesis of a functional protein. Using an intron probe derived from a partial clone of the human fibroblast ADP/ATP translocase gene, we localized the gene to chromosome Xq13----Xq25-26. The gene encoding the skeletal muscle translocase has previously been shown to be on chromosome 4. Therefore, the human ADP/ATP translocase genes are members of a multigene family that includes pseudogenes and has been dispersed to at least two chromosomes.
Asunto(s)
Cromosomas Humanos Par 10/ultraestructura , Translocasas Mitocondriales de ADP y ATP/genética , Nucleotidiltransferasas/genética , Seudogenes , Secuencia de Aminoácidos , Secuencia de Bases , Southern Blotting , Línea Celular , Mapeo Cromosómico , Cromosomas Humanos Par 10/análisis , ADN/análisis , ADN/genética , Fibroblastos/metabolismo , Expresión Génica , Humanos , Células Híbridas/citología , Células Híbridas/enzimología , Intrones , Datos de Secuencia Molecular , Mapeo Restrictivo , Transcripción GenéticaRESUMEN
The gene coding for the alpha 1 chain of human type XIII collagen. COL13A1, is assigned to chromosome region 10q11----qter by Southern blot hybridization of DNA from 24 human x rodent somatic cell hybrids using a cloned cDNA as probe. A number of previous reports indicate that 10 of the collagen genes are located on six autosomes, but no other collagen genes have been found on chromosome 10. The data therefore provide further evidence for the dispersion of members of the collagen gene family throughout the genome.
Asunto(s)
Mapeo Cromosómico , Cromosomas Humanos Par 10/ultraestructura , Colágeno/genética , Genes/genética , Animales , Southern Blotting , Cromosomas Humanos Par 10/análisis , Cricetinae , Cricetulus , ADN/análisis , ADN/genética , Sondas de ADN , Biblioteca Genómica , Humanos , Células Híbridas/citología , Células Híbridas/ultraestructura , Ratones , Hibridación de Ácido NucleicoAsunto(s)
Mapeo Cromosómico , Cromosomas Humanos Par 10/análisis , Ligamiento Genético , Marcadores Genéticos/análisis , Polimorfismo Genético , Cromosomas Humanos Par 10/ultraestructura , Enfermedades del Sistema Endocrino/genética , Humanos , Neoplasias/genética , Linaje , Polimorfismo de Longitud del Fragmento de Restricción , Recombinación GenéticaRESUMEN
The practical applications and improvements to a fast scanning microscope photometer system are described. The production of a density profile is demonstrated which, when taken in conjunction with the chromosome scan, allows more information to be extracted from the data than was previously possible. This enables a more accurate correlation to be made between phenotype and karyotype. The system is demonstrated on preparations of known cytogenetically abnormal individuals, showing the range of problems which can be tackled.