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- W2553574184 abstract "Keratinocyte Growth Factor (KGF) is a paracrine-acting and epithelium-specific growth factor produced by cells ofmesenchymal origin. Based on preclinical data, recombinant KGF plays a critical role in protecting and repairing ofdamaged epithelial tissues. Despite great efforts to express recombinant human KGF(rhKGF) in different organisms,attempts for finding appropriate protein expression system with the ability of producing a properly folded andprocessed KGF needs further investigation. Pichia pastoris has been used successfully and extensively for productionof industrial enzymes and pharmaceutical proteins. Herein, we investigated the affect of pro-region-α-factor earlydeletion on production and secretion of rhKGF in Pichia pastoris. Initially, expression of human KGF induced inMCF-7 cell line treated with 1, 25-Dihydroxy vitamin D3. The coding sequence of full-length rhKGF194 was thencloned into the yeast integrative expression vector, downstream of α-factor and was integrated into P. pastorisgenome. KGF protein was expressed in P. pastoris x33 cells, usingα-factor signal peptide for translocation of KGFto ER. An internal human signal peptide was also arranged after α-factorfor early removal of the pro-region in ER.RT-PCR results demonstrated that KGF mRNA was expressed successfully after induction by methanol.Recombinant KGF protein expression was detected by Western blotting in cell lysats, but not in conditioned media.A molecular weight of 17 kD for rhKGF194 indicates that the α-factor and internal human signal peptideshad beenremoved in x33 cells. The results indicate that in the absence of pro-region-α-factor, the recombinant KGF proteinwas not efficiently processed and transported within the biosynthesis-secretory pathway. As KGF protein is anunstable growth factor and tend to aggregate because of some native properties, It seems that presence of a chaperonmolecule fusion with KGF is necessary for efficient secretion of the recombinant protein." @default.
- W2553574184 created "2016-11-30" @default.
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- W2553574184 date "2016-07-01" @default.
- W2553574184 modified "2023-09-26" @default.
- W2553574184 title "Expression of the Full-length Human Recombinant Keratinocyte Growth Factor in Pichia pastoris" @default.
- W2553574184 doi "https://doi.org/10.22067/jcmr.v8i1.50029" @default.
- W2553574184 hasPublicationYear "2016" @default.
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