[Expression of quail vascular endothelial growth factor receptor quek-1 extracellular domain 2-4 loop cDNA in Pichia pastoris].
The extracellular domain 2-4 loop cDNA of quail vascular endothelial growth factor receptor quek1 (qVEGFR2) was obtained from plasmid carrying qVEGFR2 by PCR. Then it was cloned into expression vector pPICZalphaA of Pichia pastoris. To construct recombinant expression plasmid pPICZalphaA-qVEGFR2, linearized pPICZalphaA-qVEGFR2 with SacI was transformed to electroporated Pichia pastoris GS115. Subsequently, positive clone was selected by PCR and its phenotype was determined. SDSPAGE and Western blot assays of culture medium from a methanol-induced expression strain demonstrated that recombinant qVEGFR2 proteins were expressed and secreted into the culture medium. These results could provide a basis for further researches on tumor protein vaccine as well as for the preparation of tumor protein vaccine with Pichia pastoris.
Diao
Peng
P
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.
Wen
Yanjun
Y
Wang
Yongsheng
Y
Do
Xiaobo
X
Zhou
Hang
H
Wei
Yuquan
Y
D000818:Animals; D019496:Cancer Vaccines; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011784:Quail; D017981:Receptors, Neurotransmitter / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000096:biosynthesis* / Q000235:genetics
D019496:Cancer Vaccines; D018076:DNA, Complementary; D017981:Receptors, Neurotransmitter; D011994:Recombinant Proteins; C083131:KDR protein, Coturnix japonica; D040301:Vascular Endothelial Growth Factor Receptor-2