تاثیر غلظت های مختلف IPTG در القا بیان ژن اینترفرون بتا 1بی در اشریشیا کلی نوترکیب
محورهای موضوعی : بیوتکنولوژی و میکروبیولوژی کاربردی
کلید واژه: اشریشیا کلی, IPTG, اینترفرون بتا 1 بی, پروموتر lac,
چکیده مقاله :
Background: Lac promoter, a regulatory gene in Escherichia coli, is a prokaryotic expression system forrecombinant protein production. This promoter is induced by lactose or IPTG ,its chemical analog. In this study,the effect of different concentrations of IPTG on the expression of Interferon β-1b was investigated.Materials and Methods: Escherichia coli BL21 (DE3) harboring Interferon β-1b gene were cultured at 37°Cand induced by the addition of different concentrations of IPTG at OD600nm=0.6 , after incubation for 6 houres,the cells were collected by centrifugation and disrupted with a sonicator. A compact pellet containing inclusionbodies was obtained and applied to SDS–PAGE gel and the expression yield was analyzed using the QuantityOne quantitative software (Bio-Rad) according to the relative band intensities of Coomassie blue stain.Results:The yield of Interferon β-1b protein increased by using 1 mM IPTG as inducer and it was equal to 36%of the total cell proteins.Conclusion: The most appropriate concentration of IPTG for expression of Interferon β-1b protein is aconcentration of 1 mM.
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