Expression and detection of recombinant directing toxin SL120 in Pichia pastoris.
- Author:
Hong WANG
1
;
Ning-Yi JIN
;
Ge-Fen YIN
;
Yi-Ming XU
;
Hong-Tao JIN
;
Li-Shu ZHANG
;
Zi-Jian LI
Author Information
1. Key laboratory of Genetic Engineering of PLA, Academy of Military Medical Sciences, Changchun 130062, China.
- Publication Type:Journal Article
- MeSH:
Enterotoxins;
genetics;
Genetic Vectors;
HIV Envelope Protein gp120;
biosynthesis;
genetics;
immunology;
HIV-1;
genetics;
immunology;
Immunoglobulin Variable Region;
biosynthesis;
genetics;
immunology;
Pichia;
genetics;
metabolism;
Recombinant Proteins;
biosynthesis;
genetics;
immunology
- From:
Chinese Journal of Biotechnology
2005;21(3):473-477
- CountryChina
- Language:Chinese
-
Abstract:
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.