High level expression of interleukin-1 beta in a recombinant Escherichia coli strain for use in a controlled bioreactor.
A recombinant bacterial strain for the large scale production of human interleukin-1 beta (IL-1 beta) was constructed. The lambda Pr and the tryptophan systems were compared for efficiency of transcription and regulation. The efficiency of IL-1 protein production from these constructs was analyzed. Enhanced protein synthesis was achieved by the fusion of lambda Pr promoter sequences with trp leader sequences which included the trp RBS. A strain (JM101) was selected for use as a host and tested in a one liter bioreactor. A growth and induction regimen was established for use in bioreactors which results in the accumulation of 0.75-0.95 g l-1 of recombinant IL-1.
Otsuka Pharmaceutical Co., Ltd., Rockville, Maryland 20850.
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PM
Arthur
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Duckworth
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Seidman
D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D014364:Tryptophan / Q000235:genetics
D007375:Interleukin-1; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014364:Tryptophan; D004247:DNA