Title: Expression of recombinant human Epidermal Growth Factor (EGF) in E. coli BL21
Name: Hamidreza iranpoor
Golestan University of Medical Sciences, Gorgan, Iran
Introduction: Human Epidermal Growth Factor (HrEGF), has 53 amino acid with a molecular weight of 6.45 kDa, which is mainly synthesized by macrophages, urine, saliva, milk, and plasma. These proteins are known pharmaceutical brand “Rgen-D” and effective application inthe healing of oral and gastroesophageal ulcers, inhibition of gastric acid secretion, stimulation of DNA synthesis as well as mucosal protection from intraluminal injurious factors such as gastric acid, bile acids, pepsin, and trypsin. This study investigated the expression of therapeuticproteins in E.coli Origami to produce a recombinant form of Human Epidermal Growth Factor.
Methods: In this descriptive study, EGF gene codon optimization after Bioinformatics were synthesized and gene was cloned in the pET-24a by cutting strategy with Nde1 and BamH1-HF enzymes and purified from the gel. The constructed expression cassette was transfromed into E.coli through heat shock and CaCl2 method. Identification and confirmation of transformed colonies was performed using screening PCR. Induction of the prtoein was done with IPTG. The expression in induced strains was analyzed by SDS-PAGE and western blotting techniques.Confirmation of cloning and IGF-1 expression cassette was carried out through genetic engineering procedures.
Results: Expression analysis of transformed E.coli strain with SDS-PAGE and western blotting confirmed that gene integrated into host as well as expression. Molecular weight of expressed protein has been estimated to be 6.45 kDa.
Conclusion: EGF expression cassette for cloning and expression in E.coli was designedand protein of interest was successfully induced and identified E.coli can be used as a suitable host for production of recombinant EGF and this technology has a potential for localization.
Biography
Hamidreza Iranpoor has completed his MSc at the age of 25 years from Golestan University of Medical Sciences. He has published 1 paper in reputed journals.
Presenting author details
Full name: Hamidreza iranpoor
Contact number: 00989133051562
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Sessionname/ number: Title: Expression of recombinant human EGF (EGF) in E. coli BL21
Category: Oral presentation