Please use this identifier to cite or link to this item: https://hdl.handle.net/11147/6677
Title: Cloning, expression, and activity analysis of human cathepsin C in the yeast pichia pastoris
Authors: Dağlıoğlu, Cenk
Dağlıoğlu, Cenk
Izmir Institute of Technology. Molecular Biology and Genetics
Keywords: Expression
Human cathepsin C
Pichia pastoris
Recombinant protein
Issue Date: 2017
Publisher: TUBITAK
Source: Dağlıoğlu, C. (2017). Cloning, expression, and activity analysis of human cathepsin C in the yeast pichia pastoris. Turkish Journal of Biology, 41(5), 746-753. doi:10.3906/biy-1704-4
Abstract: The yeast Pichia pastoris expression system was investigated for the production of human cathepsin C (CatC) recombinant protein. The full-length CatC cDNA, corresponding to amino acids 12-475, was synthesized from interleukin-2 (IL-2) stimulated human peripheral blood mononuclear cells and subcloned in the pGEM-T cloning vector. After confirming the DNA sequence of the insert, the gene was cloned into the pPICZαA expression vector under the control of the methanol-inducible alcohol oxidase (AOX1) promoter and transformed to P. pastoris X-33 cells. The expressed protein was secreted into the culture medium through the α-factor mating signal sequence of the expression vector. Analysis of the culture supernatant revealed that the recombinant human CatC was secreted as a 58-kDa molecule, indicating that human CatC was accumulated in the culture supernatant as proform composed of the residual propart, the activation peptide, and the heavy and light chains. Extracellular recombinant proCatC was further activated by cysteine endoprotease papain in vitro and its activity was confirmed by assays using a synthetic substrate.
URI: http://doi.org/10.3906/biy-1704-4
http://hdl.handle.net/11147/6677
ISSN: 1300-0152
1303-6092
Appears in Collections:Molecular Biology and Genetics / Moleküler Biyoloji ve Genetik
PubMed İndeksli Yayınlar Koleksiyonu / PubMed Indexed Publications Collection
Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
TR Dizin İndeksli Yayınlar / TR Dizin Indexed Publications Collection
WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection

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