RESUMO
Double-stranded DNA encoding the human hormone somatomedin-C (SMC) has been synthesized. This synthetic gene has been inserted into a plasmid bearing the strong leftward promoter (PL) of bacteriophage lambda and expressed in E. coli. Codons for the N-terminal region of SMC which maximized the hormone's synthesis were chosen in an SMC-lac z fusion assay. The amounts of SMC accumulated in E. coli were influenced by mutations at two chromosomal loci, lon and htpR.
Assuntos
Proteínas de Escherichia coli , Escherichia coli/genética , Protease La , Serina Endopeptidases , Somatomedinas/genética , Proteases Dependentes de ATP , Clonagem Molecular , DNA Recombinante , Endopeptidases/genética , Regulação da Expressão Gênica , Genes , Genes Bacterianos , Proteínas de Choque Térmico/genética , Fator de Crescimento Insulin-Like I , Mutação , Conformação de Ácido Nucleico , Regiões Promotoras GenéticasRESUMO
Streptomyces lividans 66 was transformed with a plasmid containing the regulatory region of the Streptomyces fradiae aph gene and a structural gene that specifies bovine growth hormone (bGH). When grown in liquid culture the transformant contained a protein identical to authentic bGH, as judged by radioimmunoassay and immuno-blotting (Western analysis). The bGH was present in cells that had been in culture for up to four weeks but was not found in the medium. The strategy employed should be generally applicable to the expression of foreign genes in actinomycetes.