Basic Immunology
Expression of a novel adjuvant TFPR1 in Pichia pastoris and its identification
Xiuzhe Ning, Zhihua Kou, Weilai Sun, Qing Zhu, Yi Yang, Hongjie Qiu, Jingjing Guo, Yan Guo, Hong Yu, Yusen Zhou
Published 2016-04-30
Cite as Chin J Microbiol Immunol, 2016, 36(4): 294-299. DOI: 10.3760/cma.j.issn.0254-5101.2016.04.011
Abstract
ObjectiveTo express a recombinant protein TFPR1 (the functional region of the snake venom proteins from Trimeresurus flavoviridis) in Pichia pastoris expression system.
MethodsThe target gene was codon-optimized and synthesized according to the sequence of the conserved structural domain of triflin and then cloned into the Pichia pastoris expression vector pPICZαA to construct the recombinant expression plasmid pPICZαA-TFPR1. The recombinant plasmid pPICZαA-TFPR1 was electroporated into the yeast strain X33. The transformed strains carrying expression plasmid were screened out with Zeocin and then induced by methanol to express the recombinant protein TFPR1. ELISA was performed for the screening of positive clones. SDS-PAGE and Western blot were used for further identification of the expressed products.
ResultsThe recombinant plasmid pPICZαA-TFPR1 was successfully constructed. The recombinant protein TFPR1 was expressed in a secreted form at a molecular weight of 16×103.
ConclusionThe recombinant protein TFPR1 was successfully expressed in Pichia pastoris expression system, which laid a foundation for further researches on its biological function and application as an adjuvant.
Key words:
Adjuvant; Pathogenesis related protein 1 (PR-1); Pichia pastoris; Secretory expression
Contributor Information
Xiuzhe Ning
School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
Zhihua Kou
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China
Weilai Sun
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China
Qing Zhu
School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
Yi Yang
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China
Hongjie Qiu
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China
Jingjing Guo
School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
Yan Guo
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China
Hong Yu
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China
Yusen Zhou
School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
