RESUMO
Bovine leukocyte adhesion (BLAD) is a recessive autosomal disease in Holstein-Friesian cattle caused by point mutation in CD18 gene encoding neutrophil-surface glycoprotein. To determine BLAD carriers, the convenient primers were chosen to amplify the mutant region of gene with the following restriction analysis. A screening program for BLAD has been initiated. Among 190 animals from different Ukrainian farms 6 were heterozygous according to the tested trait, i.e., BLAD deficient. No homozygous BLAD carriers were detected.
Assuntos
Doenças dos Bovinos/genética , Síndrome da Aderência Leucocítica Deficitária/genética , Síndrome da Aderência Leucocítica Deficitária/veterinária , Mutação/genética , Alelos , Animais , Sequência de Bases , Bovinos , DNA/genética , DNA/isolamento & purificação , Triagem de Portadores Genéticos/métodos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , UcrâniaRESUMO
The method for testing foreign plasmid DNA mutagenicity on the competent culture of B. subtilis has been developed. High mutagenic effect of DNA of recombinant plasmids carrying a single human Alu-repeat or the same repeat in combination with human apoAi gene or human insulin gene was demonstrated. The vector plasmid pUC18 had no mutagenic activity. According to the data of dot-blotting some fragments of recombinant plasmid DNA of human origin can integrate in B. subtilis chromosome by means of illegitimate recombination. It is concluded that B. subtilis test system is suitable for detection of potential mutagenic polynucleotide sequences in recombinant plasmid constructions produced for gene therapy purposes.
Assuntos
Bacillus subtilis/genética , DNA Bacteriano/genética , DNA Recombinante/genética , Mutagênese , Plasmídeos/genética , Apolipoproteína A-I/genética , Cromossomos Bacterianos , Terapia Genética , Humanos , Insulina/genética , Sequências Repetitivas de Ácido Nucleico/genéticaRESUMO
The mutagenic activity of the pUC19 bacterial plasmid DNA and the pAins recombinant plasmid DNA carrying human insulin gene has been investigated. Both pUC19 and pAins plasmid DNAs have been shown to induce the gene mutations in hprt locus of Chinese hamster cell line. The high level of the gene mutations (similar to the indices of the gene mutations induced by the chemical mutagens) has been in the focus of attention. The conclusion has been made concerning impossibility to use pAins plasmid DNA in the diabetes mellitus gene therapy. It is necessary to test the mutagenic properties of the DNA molecules produced for gene therapy of human inherited diseases.
Assuntos
Genes/genética , Insulina/genética , Mercaptopurina/antagonistas & inibidores , Mutagênese/genética , Plasmídeos/genética , Recombinação Genética/genética , Animais , Células Cultivadas , Cricetinae , Cricetulus , DNA/genética , Resistência a Medicamentos/genética , Humanos , Transfecção/genéticaRESUMO
It is confirmed that inhibitors of DNA repair caffeine and 3-aminobenzamide decrease the survival of gamma-irradiated HeLa cells. It is shown that the decreased survival of irradiated cells is reversed when Escherichia coli RecA protein is introduced into cell nucleases with the aid of liposomes. This effect is more expressed in caffeine-treated (before or after irradiation) than in 3-aminobenzamide-treated (before irradiation) cells. It is suggested that E. coli 38 kD RecA protein may compensate the function of HeLa RecA-like protein, inhibited by DNA repair inhibitors, which is necessary for the repair of single-strand breaks and double-strand breaks of DNA.
