Expression line approach to recombinant human epidermal growth factor into the yeast, Pichia pastoris from Huh-7 cell line
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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.
KeywordsHuman epidermal growth factor Human hepatoma cell line Pichia pastoris Polymerase chain reaction
We thank to Molecular Medicine Lab (CEMB) for providing Huh-7 cell line. Thanks to Ahmad Usman Zafar, Nadeem Ahmad, Muhammad Islam Khan and Tayyab Hussnain for thoughtful discussions and critical reading of the manuscript.
Conflict of interest
The authors declare that they have no competing interests.
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