Bustami, Yazmin
(2010)
Cloning And Expression Of Synthetic Human
Erythropoietin In Methylotrophic Yeast, Pichia Pastoris.
Masters thesis, Universiti Sains Malaysia.
Abstract
Human erythropoietin is a hormone or glycoprotein produced by the kidney
and bone marrow with important physiological functions, such as, erythropoiesis,
angiogenesis, and wound healing. The natural source of huEpo is purified from
human urine (uhuEPO). Alternatively, recombinant human erythropoietin could
overcome the limited supply of uhuEPO. Thus, the focus of this study is to generate a
synthetic huEPO gene and express in Pichia pastoris. PCR assembly method utilized
to construct a synthetic gene using twenty sets of overlapping oligonucleotides,
covering the huEPO A gene sequence and two newly introduced restriction enzyme
sites. Results from DNA sequence analysis showed accurate assembled synthetic
huEPO gene albeit minor base mutations detected.
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