Expression of recombinant human Interleukin 2 (rhIL-2) in E. coli

  • سال انتشار: 1394
  • محل انتشار: کنفرانس بین المللی علوم و مهندسی
  • کد COI اختصاصی: ICESCON01_0029
  • زبان مقاله: انگلیسی
  • تعداد مشاهده: 872
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نویسندگان

Hamidreza Iranpoor

(MSc) Department of Medical Biotechnology, Faculty of Advanced Technologies In Medicine, Golestan University of Medical Science, Gorgan, Iran

Venus Vatankhah

(MD) Medical school of Shahrekord University of Medical Sciences, Shahrekord, Iran

چکیده

Background: interleukin-2 (IL-2) is a protein consisted of 132 amino acids (MW = 1.53 kDa). Aldesleukin is the synthetic form of the protein which is used as an effective function for immuno system. Current study was aimed to investigate the expression of rhIL-2 in E. coli BL21 (DE3) expression system in order to produce an active recombinant form of the protein. Methods: Firstly codon optimization was done for hIL-2 gene. Then the gene was synthesized and inserted in pET-22a vector by a cutting strategy included NdeI and BamHI-HF enzymes. In the next step, gene was run in agarose gel and purified. The constructed expression cassette was transformed into E. coli BL21 (DE3) cells through CaCl2 heat shock method. Identification and confirmation of the transformed colonies were performed using screening PCR method. Synthesis of rhIL-2 was induced by IPTG. The expression in induced strains was analyzed by SDS-PAGE and western blotting techniques. Results: Analysis of transformed E. coli strain with SDS-PAGE and western blotting techniques confirmed that gene was expressed in host cells. Molecular weight of the expressed protein was estimated to be 657 kDa. Conclusion: rhIL-2 expression cassette for cloning and expression in E. coli was designed and the protein of interest was successfully induced and identified. E. coli BL22 (DE3 ) can be used as a suitable host for production of recombinant hIL-2 and this technology has a potential to be localized.

کلیدواژه ها

Interleukin 2, E. coli, Gene expression, cloning

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