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1.
Mol Biol (Mosk) ; 25(2): 431-41, 1991.
Artigo em Russo | MEDLINE | ID: mdl-1679193

RESUMO

Expression and structural organization of tyrosine aminotransferase (TAT) gene in Morris hepatoma cell line 7777 with active and glucocorticoid-inducible TAT gene and in hepatoma 8994, where TAT gene does not function were analysed. No differences in the number of receptor macromolecules, translocation and nuclear binding of hormone-receptor complexes in hormone sensitive (7777) and resistant (8994) cell lines were demonstrated. Dexamethasone increases TAT gene transcription in 7777 cell line but not 8994. Restriction analysis of TAT gene does not reveal any differences either in structural or in regulatory regions. Gel retardation assay with cloned TAT fragment (-400 b.p.) from normal hepatocytes showed identical shift of mobility in 7777 and 8994 cell lines. Moreover, 5'-flanking sequence (-890 b.p.) of TAT gene linked to the bacterial CAT gene is transiently expressed in both cell lines. We have shown that HpaII site (-105 b.p.) of TAT gene is methylated in those cells where TAT gene does not function (thymus, spleen, Zajdela ascites hepatoma) and is demethylated in TAT gene expressing hepatoma 7777 and normal rat hepatocytes. In hepatoma 8994 there are no DNAse I hypersensitive regions, typical to functioning TAT gene from hepatoma 7777 and normal hepatocytes.


Assuntos
Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas Experimentais/enzimologia , Tirosina Transaminase/genética , Animais , Cloranfenicol O-Acetiltransferase/genética , DNA de Neoplasias/genética , DNA de Neoplasias/metabolismo , Genes tat , Metilação , Plasmídeos , RNA Mensageiro/genética , Ratos , Mapeamento por Restrição , Transcrição Gênica , Transfecção
2.
Mol Biol (Mosk) ; 24(1): 163-72, 1990.
Artigo em Russo | MEDLINE | ID: mdl-2190080

RESUMO

We have studied the effect of some regular sequences namely (dA-dT)n, (dA)n and (dG-dC)n on the Saccharomyces cerevisiae PHO5 gene expression. These sequences were inserted into the ClaI site, located between two phosphate-responsible upstream activating sequences (UAS's). A modified PHO5 gene cloned in vector plasmids was used to transform the pho3, pho5 yeast strain and the level of acid phosphatase expression in various conditions was measured. We show that the insertion of (dA-dT)n blocks, but not (dA)n or (dG-dC)n, leads to the dramatic decrease of PHO5 gene expression in a derepressed state. (dA-dT)n inserts also inhibit the expression of PHO5 gene which was put under the control of heterologous UASgal.


Assuntos
Regulação Fúngica da Expressão Gênica , Genes Fúngicos , Sequências Reguladoras de Ácido Nucleico , Saccharomyces cerevisiae/genética , Fosfatase Alcalina/genética , Desoxirribonuclease EcoRI , Dados de Sequência Molecular , Plasmídeos , Transcrição Gênica
3.
Mol Gen Mikrobiol Virusol ; (6): 42-5, 1987 Jun.
Artigo em Russo | MEDLINE | ID: mdl-3041203

RESUMO

A chromosomal gene for human leucocyte interferon A is expressed in Saccharomyces cerevisiae yeasts due to interaction of 5'-nontranslating region of the cloned interferon gene with the regulatory elements of yeast genes PHO5, GAL1 and GAL10. Regulated systhesis of interferon was obtained in all cases. The level of interferon genes expression in case using GAL1 and GAL10 genes regulatory elements (5 X 10(5) and 5 X 10(6) u X l-1) correlated with the distances to their promoters. The highest yield of interferon (10(8) u X l-1) was obtained when the PHO5 gene regulatory elements were used.


Assuntos
Regulação da Expressão Gênica , Genes Fúngicos , Genes Reguladores , Interferon Tipo I/genética , Saccharomyces cerevisiae/genética , Clonagem Molecular , Enzimas de Restrição do DNA , Humanos , Plasmídeos , Transformação Genética
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