RESUMEN
A modified system with two pairs of primers is suggested for the analysis of different bcr/abl mRNA variants, using the polymerase chain reaction (PCR) method. It was demonstrated that, with the use of this method, mRNA preparations obtained from bone marrow cells are more informative than those obtained from blood cells. Fresh preparations of thermostable Tth DNA polymerase, which exhibited the reverse transcriptase activity, appeared to be optimal for PCR analysis of the bcr/abl mRNA pattern.
Asunto(s)
Proteínas de Fusión bcr-abl/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , ARN Mensajero/metabolismo , Secuencia de Bases , Cartilla de ADN , ADN Polimerasa Dirigida por ADN/metabolismo , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ADN Polimerasa Dirigida por ARN/metabolismoRESUMEN
The site of the break of chromosome 22 was investigated by blot hybridization and polymerase chain reaction in 41 patients with chronic myeloid leukemia (CML). It is stated that the break site in various regions is not associated with the disease stage and clinicohematological manifestations in the time of the diagnosis. The break in the 5' subregion was indicative of longer survival than that in the subregion 3' patients. Median survival reached 60.8 and 32.4 months, respectively.
Asunto(s)
Cromosomas Humanos Par 22 , Cromosomas Humanos Par 9 , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Translocación Genética , Adolescente , Adulto , Anciano , ADN de Neoplasias/genética , Femenino , Humanos , Leucemia Mielógena Crónica BCR-ABL Positiva/mortalidad , Masculino , Persona de Mediana Edad , Cromosoma Filadelfia , Pronóstico , ARN Neoplásico/genéticaRESUMEN
Using oligonucleotide probes, sequences containing the Mbcr locus involved in chromosome translocation t(9:22) were cloned form the library of human genes in the Charon 4A vector. The recombinant clone lambda BCR 1.1 obtained contained Mbcr sequences, but the 3' region of the Mbcr locus in lambda BCR 1.1 clone was strongly altered. Subcloning of a fragment of the altered region and blot hybridization analysis using it as a DNA probe revealed recombination in the 3' region of the Mbcr locus in clone lambda BCR 1.1 which resulted in insertion of unknown sequences into the region. A modified system is suggested for chromosome 22 breakpoint identification using restriction analysis of genome DNA with four restriction endonucleases and one 5'-DNA probe.
Asunto(s)
Cromosomas Humanos Par 22 , Cromosomas Humanos Par 9 , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Translocación Genética , Secuencia de Bases , Clonación Molecular , Humanos , Datos de Secuencia Molecular , Sondas de Oligonucleótidos , Recombinación Genética , Mapeo RestrictivoRESUMEN
Using as a hybridization probe cDNA 35s RNA from the Rauscher leukemia cells, a part of rRNA gene cluster from the gene library of mice C-1 erythroleukemia cells has been cloned. Fragment 6.7 kb recloned into pUC19 rRNA was used as a probe to analyse organization of rRNA genes of mice with RL. The observed amplification and rearrangement in genome DNA of spleen cells are determined by a new type of their rRNA genes rearrangement in the nontranscribed as well as in the transcribed part of rRNA.