RESUMEN
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.
Asunto(s)
Amplificación de Genes , Reordenamiento Génico , Genes , Leucemia Experimental/genética , ARN Ribosómico/genética , Animales , Clonación Molecular/métodos , ADN , Sondas de ADN , Leucemia Eritroblástica Aguda/genética , Ratones , Ratones Endogámicos AKR , Ratones Endogámicos BALB C , Hibridación de Ácido Nucleico , Radioisótopos de Fósforo , Poli A , Polimorfismo de Longitud del Fragmento de Restricción , Sondas ARN , Virus RauscherRESUMEN
The restriction analysis of BALB/c and AKR mice genome DNA was used to show the translocation and amplification of gene which is expressed with the Rauscher erythroleukemia as the 35S nuclear RNA. The homology of this RNA to 10 defined oncogenes was studied. It was found that the cDNA 35S RNA is efficiently hybridized only with v-sis oncogene. The clones homologous to the 35S RNA, v-sis oncogene and 3'-area of the Rauscher leukemia virus genome are isolated from the genes' library of mice erythroleukemic cells.
Asunto(s)
Secuencia de Bases , ADN de Neoplasias/genética , Leucemia Experimental/genética , Proto-Oncogenes , Homología de Secuencia de Ácido Nucleico , Bazo/análisis , Animales , Enzimas de Restricción del ADN , ADN de Neoplasias/análisis , Leucemia Eritroblástica Aguda/genética , Ratones , Ratones Endogámicos BALB C , Hibridación de Ácido Nucleico , Virus RauscherRESUMEN
The library of genes was obtained from erythroleukemic AKR cells (C-1), that were maintained as suspension culture. Thirty four clones that had homology with 60-70S RNA of Rauscher Leukemia virus (RLV) were separated from this library. The restriction mapping was carried out with 14 clones, that contained most extensive proviral sequences. One clone (107) contains proviral sequences that are derived from one of the components of the RLV complex. The other 13 clones contain sequences of endogenous xenotropic viruses. The endogenous retroviral sequences obtained differ in restrictive maps from proviruses of ecotropic and xenotropic infectious endogenous MuLV and, apparently, might be attributed as non-inducible infectious xenotropic MuLV of class III. Some of the cloned retroviral sequences had symmetrical structure, that is typical for integrated proviruses, i. e. these sequences were separated from flanking cellular ones by long terminal repeats. All investigated retroviral sequences are deletion mutants of MuLV proviruses. It was shown that the inner regions of proviruses diverged more than the long terminal repeats. The expression of the main inner MuLV polypeptide (p30) was detected in NIH 3T3 cells, transfected with DNA of some clones.
Asunto(s)
Clonación Molecular , ADN Viral/genética , Virus de la Leucemia Murina/genética , Leucemia Eritroblástica Aguda/microbiología , Recombinación Genética , Animales , Línea Celular , Mapeo Cromosómico , ADN/genética , Enzimas de Restricción del ADN , ADN de Neoplasias/genética , Leucemia Eritroblástica Aguda/genética , Ratones , Ratones Endogámicos AKR , Plásmidos , Virus Rauscher/genéticaRESUMEN
The pigment-protein complexes enriched with photosystem I (PPC-1) and photosystem II (PPC-2) were obtained using sievorptive chromatography on DEAE-Sephadex. Both types of complexes contain chlorophyll a, beta-carotene and minor quantities of chlorophyll b. Red absorption maxima for PPC-1 and PPC-2 are located at 676 and 673 nm, respectively. Using differential spectrophotometry, the degree of the reaction centre enrichment was established: PPC-1 has one P700 per 35 bulk chlorophyll a molecules, PPC-2 contains one P680 per 18 bulk chlorophyll a molecules. The yield of PPC-2 is 7-10 times lower than that of PPC-1 and equals to 0,3% of the chlorophyll content of the initial chloroplasts. The amount of P680 in PPC-1 preparations does not exceed 7% after a single chromatographic procedure; the amount of P700 in PPC-2 makes up to 2%. The method proposed is more advantageous with respect to higher reaction centre enrichment of PPC-1 and PPC-2 as compared to ion-exchange chromatography on DEAE-cellulose.