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The regulation of protein tyrosine phosphorylation is mediated by protein tyrosine kinases(PTKs) and protein tyrosine phosphatases (PTPs) and is essential for cellular homeostasis. Coexpressionof PTKs with PTPs in Pichia pastoris was used to facilitate the expression of activePTKs by neutralizing their apparent toxicity to cells. In this study, the gene encodingphosphatase PTP1B with or without a blue fluorescent protein or peroxisomal targeting signal1 was cloned into the expression vector pAG32 to produce four vectors. These vectors weresubsequently transformed into P. pastoris GS115. The tyrosine kinases EGFR-2 and PDGFRβwere expressed from vector pPIC3.5K and were fused with a His-tag and green fluorescentprotein at the N-terminus. The two plasmids were transformed into P. pastoris with or withoutPTP1B, resulting in 10 strains. The EGFR-2 and PDGFRβ fusion proteins were purified by Ni2+affinity chromatography. In the recombinant P. pastoris, the PTKs co-expressed with PTP1Bexhibited higher kinase catalytic activity than did those expressing the PTKs alone. Thehighest activities were achieved by targeting the PTKs and PTP1B into peroxisomes. Therefore, the EGFR-2 and PDGFRβ fusion proteins expressed in P. pastoris may be attractivedrug screening targets for anticancer therapeutics.

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