title: Expression line approach to recombinant human epidermal growth factor into the yeast, Pichia pastoris from Huh-7 cell line.
abstract: Beta-urogastrone also known as human epidermal growth factor is a key member of epidermal growth factor family having role in cell proliferation and differentiation in vivo as well as in vitro. Human epidermal growth factor gene has been isolated from different tissues but the method of isolation is technically difficult and complicated as it deals with biopsies. Here we isolated mature partial human epidermal growth factor gene from Huh-7 cell line, amplified and abridged toward mature coding region with three steps PCR, sequenced for homology with wild type human epidermal growth factor gene, inbuilt with sites of interest and cloned in Pichia pastoris for expression study. Isolated mature human epidermal growth factor gene from Huh-7 cell line showed 100 % sequence homology to wild type human epidermal growth factor gene and gives the native expression for human epidermal growth factor peptide. In this study we report that Huh-7 cell line is an easy source for the particular gene of human epidermal growth factor isolation and we are also suggesting P. pastoris is an expression system to produce recombinant human epidermal growth factor of the therapeutic importance resembling to the natural human system.
authors.affiliation: National Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road, Thokar Niaz Baig, Lahore, Pakistan.
authors.forename: Mohsin Ahmad
authors.initials: MA
authors.lastname: Khan
authors.forename: Faidad
authors.initials: F
authors.lastname: Khan
authors.forename: Nadeem
authors.initials: N
authors.lastname: Ahmad
authors.forename: Muhammad Islam
authors.initials: MI
authors.lastname: Khan
authors.forename: Ahmad Usman
authors.initials: AU
authors.lastname: Zafar
authors.forename: Tayyab
authors.initials: T
authors.lastname: Husnain
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D049109:Cell Proliferation; D004815:Epidermal Growth Factor / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D004815:Epidermal Growth Factor