Assuntos
Bradicinina/genética , Genes Sintéticos , Vetores Genéticos , Vírus 40 dos Símios/genética , Animais , Sequência de Bases , Bradicinina/fisiologia , Células Cultivadas , Chlorocebus aethiops , DNA Recombinante , Genes Virais , Dados de Sequência Molecular , Contração Miocárdica , Plasmídeos , Ratos , Proteínas do Envelope Viral/genéticaRESUMO
The purpose of the studies is to design the recombinant virus SV 40 where the C-end of the basic structural protein of virus P-1 was replaced by a synthetic sequence of the neuropeptide bradykinin. The recombinant virus SV (SV 40/Brd) was obtained. In this virus 60 n.p. with 3'-end of VP-1 gene was substituted for 36 n.p. synthetic gene of bradykinin without impairing the frame of translation. The biological activity of SV 40 (Brd) was tested on the cultured cells CV-1 permissive for this virus. An immunofluorescence method was used to detect T antigen and to examine the cytopathic action of this recombinant. The gene engineering design does not make the recombinant loose its biological properties typical of wild virus.
Assuntos
DNA Viral/genética , Regulação Viral da Expressão Gênica/genética , Vetores Genéticos/genética , Vírus 40 dos Símios/genética , Animais , Sequência de Bases , Bradicinina/genética , Células Cultivadas/microbiologia , Efeito Citopatogênico Viral , Escherichia coli/genética , Genes Virais/genética , Dados de Sequência Molecular , Oligonucleotídeos/genética , Plasmídeos/genética , Recombinação Genética , Vírus 40 dos Símios/patogenicidade , Transfecção/genética , Proteínas Estruturais Virais/genéticaRESUMO
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 GenéticaRESUMO
Expression of a chromosomal gene for human leucocyte interferon A was obtained due to interaction of 5'-nontranslating region of a cloned interferon gene with the regulatory sequences from UNA3 yeast gene. The sequence of 5'-nontranslating region from interferon gene essential for its expression in yeast is localized within 130 b p. from the initiating codon. Due to increasing of plasmid stability and copy number a 60-fold increase. in interferon yield was obtained in yeast transformants reaching the level of 6 X 10(7) u X 1(-1). The data are presented supposing the existence of functional polycistronic mRNA in yeast.