Intracellular expression of hIFNα genes in Escherichia coli and Bacillus subtilis directed by staphylokinase signals

✍ Scribed by Reinhard Breitling; Klaus Gase; Dr. Detlev Behnke

• Publisher: John Wiley and Sons
• Year: 1990
• Tongue: English
• Weight: 390 KB
• Volume: 30
• Category: Article
• ISSN: 0233-111X
• DOI: 10.1002/jobm.3620300908

✦ Synopsis

Portable expression units for intracellular formation of heterologous proteins in Escheiichiu coli and Bacillirs subrilis were constructed by inserting the transcription and translation initiation signals of the staphylokinase salt42D gene into the polylinker of plasmid pUC18. Fusions with ATG-gene cassettes coding for mature human interferons (hIFN) c1, and x2 resulted in intracellular expression of both proteins in E. mli. The 20 fold lower yield of hIFNr, was not due to unfavorable mRNA secondary structure formation as ruled out by constructing a hybrid hIFNa,/az gene. Intracellular expression of IFNa, in B. subiilis reached 6 x I@ IU/ml. Nuclease SI mapping of transcriptional start sites revealed differential promotor usage in E. coli and B . suh/ilis. In E. coli transcription from the scik42D promoter was drastically reduced by by transcription initiating from upstream lac and ref promoters. In contrast. in B. srtbtilis transcription proceeded exclusively from the .sak42D promoler.