[Expression and characterization of human vascular endothelial growth factor receptor Flt-1 extracellular domain in Pichia pastoris].
By inserting VEGF receptor Flt-1(1-3 loop) cDNA into Pichia pastoris expression vector pPIC9K containing AOX1 promotor and the sequences of alpha secreting signal peptides, the expression plasmid pPIC9K/Flt-1(1-3) was constructed and transformed into GS115. The multi-copy insert transformants were selected and cultivated in flasks. After 4 days of 1% methanol induction, the expressed Flt-1(1-3) accumulated up to 30% of total proteins in supernatant. The expressed Flt-1(1-3) was further proved with good antigenicity and high specificity by ELISA and Western blot. They can bind to VEGF and inhibit HUVEC proliferation stimulated by VEGF.
Ma
L
L
Institute of Virology, Chinese Academy of Preventive Medicine, Beijing.
Zhang
Z Q
ZQ
Zhou
X M
XM
Zeng
G F
GF
Chen
A J
AJ
Yao
L H
LH
Wang
X N
XN
D006801:Humans; D010843:Pichia / Q000235:genetics*; D010957:Plasmids; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis* / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D040281:Vascular Endothelial Growth Factor Receptor-1
D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; D020794:Receptor Protein-Tyrosine Kinases; D040281:Vascular Endothelial Growth Factor Receptor-1