OPTIMIZATION OF CULTURAL CONDITION TO IMPROVE RECOMBINANT C-D FRAGMENT OF BACTERIORHODOPSIN PRODUCTION IN E. COLI

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KHANCHERZAR SIRVAN; HASHEMI NAJAFABADI SAMEEREH; MOHAMMADIAN MOOSA ABADI JAAFAR; KHALILZADEH RASOUL; ESFANDIAR SAMANEH

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Journal: Nashrieh Shimi va Mohandesi Shimi Iran Year:2013 | Volume:32 | Issue:2 (68) Page(s): 93-101

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Title

OPTIMIZATION OF CULTURAL CONDITION TO IMPROVE RECOMBINANT C-D FRAGMENT OF BACTERIORHODOPSIN PRODUCTION IN E. COLI

Author(s)

Keywords

E. COLIQ2

BACTERIRHODOPSINQ2

C-D FRAGMENQ2

OPTIMIZATIONQ2

RECOMBINANT PROTEINQ2

Abstract

At present study, a C-D FRAGMENt of bacteriorhodopsin (BR) in recombinant E. COLI was expressed. BR mutant gene was synthesized by consideration of E. COLI codon usage. The synthesized gene was cloned in pET21a+ expression plasmid at Nde J and Hind 111 restriction sites and expressed under T7 promoter successfully. The expressed protein was analyzed by SDS-PAGE. The effect of temperature (A), induction time (B) and the process time after induction (C), on the protein expression yield (the amount of product per unit dry cell mass) were screened using Yates table analysis. Three factors of A, Band C were significant and optimized by Taguchi method. The optimized condition was as: temperature 37oC, induction time at OD600=0.7 and the process time after induction 4 h. Also, the predicted protein production at optimized condition was 21.54% of total protein. Using the gained optimum condition, the effect of amino acid addition on expression level of recombinant C-D FRAGMENt of BR and bacterial growth in E. COLI, using M9 culture medium was also investigated. Yates table analysis showed that Threonine, Leucine and Alanine are more effective and so were selected for addition to the culture medium in the fermentor, at two different levels. The results showed that amino acid addition caused increasing in the desired protein productivity, but had little effect on the bacterial growth.

Cites

INVESTIGATION ON ACTING MECHANISM OF NITROGEN SOURCE IN CULTURE ON MEDIUM PH AND PRODUCTION OF POLY-GAMMA-GLUTAMATE BYFLAVOBACTERIUM SP.

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APA: Copy

KHANCHERZAR, SIRVAN, HASHEMI NAJAFABADI, SAMEEREH, MOHAMMADIAN MOOSA ABADI, JAAFAR, KHALILZADEH, RASOUL, & ESFANDIAR, SAMANEH. (2013). OPTIMIZATION OF CULTURAL CONDITION TO IMPROVE RECOMBINANT C-D FRAGMENT OF BACTERIORHODOPSIN PRODUCTION IN E. COLI. NASHRIEH SHIMI VA MOHANDESI SHIMI IRAN (PERSIAN), 32(2 (68)), 93-101. SID. https://sid.ir/paper/26175/en

Vancouver: Copy

KHANCHERZAR SIRVAN, HASHEMI NAJAFABADI SAMEEREH, MOHAMMADIAN MOOSA ABADI JAAFAR, KHALILZADEH RASOUL, ESFANDIAR SAMANEH. OPTIMIZATION OF CULTURAL CONDITION TO IMPROVE RECOMBINANT C-D FRAGMENT OF BACTERIORHODOPSIN PRODUCTION IN E. COLI. NASHRIEH SHIMI VA MOHANDESI SHIMI IRAN (PERSIAN)[Internet]. 2013;32(2 (68)):93-101. Available from: https://sid.ir/paper/26175/en

IEEE: Copy

SIRVAN KHANCHERZAR, SAMEEREH HASHEMI NAJAFABADI, JAAFAR MOHAMMADIAN MOOSA ABADI, RASOUL KHALILZADEH, and SAMANEH ESFANDIAR, “OPTIMIZATION OF CULTURAL CONDITION TO IMPROVE RECOMBINANT C-D FRAGMENT OF BACTERIORHODOPSIN PRODUCTION IN E. COLI,” NASHRIEH SHIMI VA MOHANDESI SHIMI IRAN (PERSIAN), vol. 32, no. 2 (68), pp. 93–101, 2013, [Online]. Available: https://sid.ir/paper/26175/en

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