EXPRESSION OF RECOMBINANT HUMAN INTERLEUKIN 2 (RHIL-2) IN E. COLI

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IRANPOOR HAMIDREZA; VATANKHAH VENUS

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Seminar Paper

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Seminar: INTERNATIONAL CONFERENCE ON SCIENCE AND ENGINEERING
Year:2015 | Issue:1

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Title

EXPRESSION OF RECOMBINANT HUMAN INTERLEUKIN 2 (RHIL-2) IN E. COLI

Author(s)

IRANPOOR HAMIDREZA | VATANKHAH VENUS

Keywords

INTERLEUKIN 2

E. COLI

GENE EXPRESSION

CLONING

Abstract

BACKGROUND: INTERLEUKIN-2 (IL-2) IS A PROTEIN CONSISTED OF 132 AMINO ACIDS (MW= 15.3 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-24A 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 7.6 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 BL21 (DE3) CAN BE USED AS A SUITABLE HOST FOR PRODUCTION OF RECOMBINANT HIL-2 AND THIS TECHNOLOGY HAS A POTENTIAL TO BE LOCALIZED.

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APA: Copy

IRANPOOR, HAMIDREZA, & VATANKHAH, VENUS. (2015). EXPRESSION OF RECOMBINANT HUMAN INTERLEUKIN 2 (RHIL-2) IN E. COLI. INTERNATIONAL CONFERENCE ON SCIENCE AND ENGINEERING. SID. https://sid.ir/paper/909096/en

Vancouver: Copy

IRANPOOR HAMIDREZA, VATANKHAH VENUS. EXPRESSION OF RECOMBINANT HUMAN INTERLEUKIN 2 (RHIL-2) IN E. COLI. 2015. Available from: https://sid.ir/paper/909096/en

IEEE: Copy

HAMIDREZA IRANPOOR, and VENUS VATANKHAH, “EXPRESSION OF RECOMBINANT HUMAN INTERLEUKIN 2 (RHIL-2) IN E. COLI,” presented at the INTERNATIONAL CONFERENCE ON SCIENCE AND ENGINEERING. 2015, [Online]. Available: https://sid.ir/paper/909096/en

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