the effect of heat shock on production of recombinant human interferon alpha 2a (rhifn α -2a) by escherichia coli
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abstract
recombinant human interferon alpha 2a (rhifn α -2a) production and cell growth were monitored in a set of genetically modified e. coli strains (msd1519, msd1520, msd 1521, msd 1522, msd 1523) producing rhifn α -2a. the growth was followed at od 600 nm, changes in cell physiology were detected by pyrolysis mass spectrometry (pyms) of cell biomass and recombinant protein production was determined by sds-gel electrophoresis. the heat stress applied was minimal (50 ° c for 5 minutes) but the effects were detected in most of the strains. all the strains except msd 1520 showed a significant increase in the quantity of the rhifn α -2a secretion at 25 hour growth under the heat shock condition, quantitated by the bio-rad molecular analyst software, multianalyst from the digital image of gels captured using a fluor s image analysis system. in the main fermentation system at t7 hour, only msd 1523 showed an increase in the rhifn α -2a secretion under the heat shock condition, at t8 hour msd 1520 and msd 1523 had an increased rhifn α -2a secretion, and at t10 msd 1521 and msd 1522 had an increased rhifn α - 2a secretion under the heat shock condition. the pccv ordination diagrams obtained from the pyms result showed, a considerable effect of heat shock on the msd 1519 strain at t5 hour. for the other strains, the result largely agreed with both the growth curves and the rhifn α -2a production that had a limited effect on e. coli cultures. the increase of temperature in the main fermentation during the log phase of the bacterial culture during rhifn α -2a expression depends on the strain specificity. this situation could definitely lead to over expression of the gene and higher intracellular accumulation of rhifn α -2a molecule. iran. biomed. j. 9 (4): 155-162, 2005
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Journal title:
iranian biomedical journalجلد ۹، شماره ۴، صفحات ۱۵۵-۱۶۲
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