[Expression and detection of recombinant directing toxin SL120 in Pichia pastoris].
Anti-HIV-1 gp120 single chain antibody(scFv) gene and staphylococcus extoxin A(SEA) gene were inserted into vector pPIC9K. The recombinant plasmid was integrated into Pichia pastoris by electroporation. High level expression was performed by determining the Muts phenotype and screening muti-copy integrants. The recombinant protein was about 57kD and the production was 50.1 mg/L. It was shown that the two kinds of protein affected the conformation of each other by antibody affinity assay, but the recombinant targeting toxins could highly mediate CTLs to kill HIV-1 target cells.
Wang
Hong
H
Key laboratory of Genetic Engineering of PLA, Academy of Military Medical Sciences, Changchun 130062, China.
Jin
Ning-Yi
NY
Yin
Ge-Fen
GF
Xu
Yi-Ming
YM
Jin
Hong-Tao
HT
Zhang
Li-Shu
LS
Li
Zi-Jian
ZJ
D004768:Enterotoxins / Q000235:genetics; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
D004768:Enterotoxins; D015699:HIV Envelope Protein gp120; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C026066:enterotoxin A, Staphylococcal