Heterologous Protein Expression in Pichia

Escherichia coli strain TOP10 was used for recombinant DNA manipulation. Pichia pastoris GS115 (Invitrogen) expressing eGFP or Phy was used as the wild-type (wt) strain and all deletion strains used in this study were derived from it. The pPICZαA (Invitrogen) vector including AOX1 promoter was used to express the heterologous protein of eGFP and Phy from Citrobacter amalonaticus CGMCC 1696 [57 (link)]. The pGAPZA (Invitrogen) vector containing GAP promoter was used to construct the RP complementation strains.

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Liao X., Zhao J., Liang S., Jin J., Li C., Xiao R., Li L., Guo M., Zhang G, & Lin Y. (2019). Enhancing co-translational folding of heterologous protein by deleting non-essential ribosomal proteins in Pichia pastoris. Biotechnology for Biofuels, 12, 38.

Publication 2019

Pichia pastoris GS115 pPICZαA pGAPZA

Citrobacter amalonaticus Deletion Egfp protein Escherichia coli Pichia pastoris Recombinant dna Strains Vector

Corresponding Organization :

Other organizations : South China University of Technology, Jinan University, East China University of Science and Technology

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Variable analysis

independent variables

Escherichia coli strain TOP10 for recombinant DNA manipulation
Pichia pastoris GS115 (Invitrogen) expressing eGFP or Phy as the wild-type (wt) strain and all deletion strains used in this study
PPICZαA (Invitrogen) vector including AOX1 promoter to express the heterologous protein of eGFP and Phy from Citrobacter amalonaticus CGMCC 1696
PGAPZA (Invitrogen) vector containing GAP promoter to construct the RP complementation strains

dependent variables

Measured outcomes of the expressed heterologous proteins (eGFP and Phy) in Pichia pastoris GS115

control variables

Pichia pastoris GS115 (Invitrogen) expressing eGFP or Phy as the wild-type (wt) strain

controls

Positive control: Pichia pastoris GS115 (Invitrogen) expressing eGFP or Phy as the wild-type (wt) strain
Negative control: Not explicitly mentioned

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