RESUMO
Chemico-enzymatic synthesis and cloning of genes coding viral proteins--precursors of p50 and gp120 in the eukaryotic vector system were carried out. The recombinant proteins obtained were shown to be used in immunochemical evaluation of AIDS virus antibodies in blood serum.
Assuntos
Western Blotting/métodos , Proteínas Recombinantes , Proteínas Virais , Síndrome da Imunodeficiência Adquirida/diagnóstico , Clonagem Molecular , Produtos do Gene gag/genética , Anticorpos Anti-HIV/análise , Antígenos HIV/genética , Proteína do Núcleo p24 do HIV/genética , Proteína gp120 do Envelope de HIV/genética , Humanos , Precursores de Proteínas/genética , Proteínas Recombinantes/síntese química , Produtos do Gene gag do Vírus da Imunodeficiência HumanaRESUMO
Chemical-enzymatic synthesis of an artificial gene encoding leader peptide and 22 N-terminal amino acids of mature carboxypeptidase G2 from Pseudomonas sp. followed by enterokinase signal sequence (Asp4Lys) has been accomplished. The resulted DNA was fused with semi-synthetic gene coding for polypeptide 4-157 of mature human tumour necrosis factor (TNF) and then placed under control of early promoters of T7 bacteriophage. The expression products of the construct obtained was analysed using anti-TNF anti-serum. In E.coli leader peptide was cleaved off during translocation through inner membrane and the resultant product was found in membrane fraction.
