Optimization of fermentation conditions for the expression of interleukin-3 in fusion with pelb in E. coli


Authors

  • Duong Thu Huong Institute of Biotechnology
  • Nguyen Thi Quy Institute of Biotechnology
  • Dang Thi Ngoc Ha Institute of Biotechnology
  • Le Thi Thu Hong Institute of Biotechnology
  • Truong Nam Hai Institute of Biotechnology
DOI: https://doi.org/10.15625/0866-7160/v38n2.7126

Keywords:

Escherichia coli JM109, IL-3, PelB, protein expression.

Abstract

Interleukin-3 (IL-3) is a multifunctional cytokine which modulates the proliferation, differentiation and maturation of various types of hematopoietic cells. Gene coding for IL-3 linked with pelB signal were incorporated into pET22b(+) for  expression of il3 gene in E. coli BL21; but IL-3 was synthesized at very low levels and still in fusion with PelB. To facilitate purification process, finding the appropriate fermentation conditions plays a key role in order to enhance gene il-3 expression and cleavage of PelB. In this study, we have optimized the conditions for the expression of IL-3, which included E. coli host strain JM109, LB cultivation medium, induction temperature was 25oC; induction with 0.05 mM IPTG at OD600 = 1. The cell biomass increases at about 2.3 times after optimization. IL-3 protein was expressed in the form of inclusion body and the PelB signal was cleaved. This result is conducive for purification of large amount of IL-3 to determine characteristics of protein.

 

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Author Biographies

Duong Thu Huong, Institute of Biotechnology

Engineering Genetic Laboratory

Nguyen Thi Quy, Institute of Biotechnology

Engineering Genetic Laboratory

Dang Thi Ngoc Ha, Institute of Biotechnology

Engineering Genetic Laboratory

Le Thi Thu Hong, Institute of Biotechnology

Engineering Genetic Laboratory

Truong Nam Hai, Institute of Biotechnology

Engineering Genetic Laboratory

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Published

26-04-2016

How to Cite

Huong, D. T., Quy, N. T., Ngoc Ha, D. T., Thu Hong, L. T., & Hai, T. N. (2016). Optimization of fermentation conditions for the expression of interleukin-3 in fusion with pelb in E. coli. Academia Journal of Biology, 38(2), 250–256. https://doi.org/10.15625/0866-7160/v38n2.7126

Issue

Vol. 38 No. 2 (2016)

Section

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