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Intracellular expression of hIFNα genes in Escherichia coli and Bacillus subtilis directed by staphylokinase signals
Author(s) -
Breitling Reinhard,
Gase Klaus,
Behnke Detlev
Publication year - 1990
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.3620300908
Subject(s) - bacillus subtilis , staphylokinase , transcription (linguistics) , escherichia coli , biology , microbiology and biotechnology , promoter , gene , plasmid , heterologous , gene expression , multiple cloning site , intracellular , expression vector , recombinant dna , genetics , bacteria , linguistics , philosophy
Portable expression units for intracellular formation of heterologous proteins in Escherichia coli and Bacillus subtilis were constructed by inserting the transcription and translation initiation signals of the staphylokinase sak 42D gene into the polylinker of plasmid pUC18. Fusions with ATG‐gene cassettes coding for mature human interferons (hIFN) α 1 and α 2 resulted in intracellular expression of both proteins in E. coli. The 20 fold lower yield of hIFNα 2 was not due to unfavorable mRNA secondary structure formation as ruled out by constructing a hybrid hIFNα 1 /α 2 gene. Intracellular expression of IFNα 1 in B. subtilis reached 6 x 10 4 IU/ml. Nuclease SI mapping of transcriptional start sites revealed differential promoter usage in E. coli and B. subtilis. In E. coli transcription from the sak 42D promoter was drastically reduced by by transcription initiating from upstream lac and tet promoters. In contrast, in B. subtilis transcription proceeded exclusively from the sak 42D promoter.