Volume 9, Issue 4 (10-2005)                   ibj 2005, 9(4): 155-162 | Back to browse issues page

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Roy R K, Sapatnekar S M, Deshmukh R A. The Effect of Heat Shock on Production of Recombinant Human Interferon Alpha 2a (rhIFN α -2a) by Escherichia coli . ibj. 2005; 9 (4) :155-162
URL: http://ibj.pasteur.ac.ir/article-1-382-en.html
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
Type of Study: Full Length |

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