Cloning and expression of single-chain Fv antibodies against H5N1 Avian influenza virus hemagglutinin.
- Author:
Shu-mei HAO
1
;
Qun LI
;
Cheng-yu WANG
;
Hua-lei WANG
;
Song-tao YANG
;
Xian-zhu XIA
Author Information
1. Changchun University of Science and Technology, Changchun 130022, China. Haosm@sina.com
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies;
genetics;
metabolism;
pharmacology;
Birds;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Gene Expression Regulation;
Hemagglutinins;
immunology;
Immunoglobulin Heavy Chains;
genetics;
Immunoglobulin Light Chains;
genetics;
Immunoglobulin Variable Region;
genetics;
metabolism;
Influenza A Virus, H5N1 Subtype;
drug effects;
immunology;
Influenza in Birds;
virology;
Mice;
Mice, Inbred BALB C;
Recombinant Fusion Proteins;
genetics;
metabolism;
pharmacology;
Reverse Transcriptase Polymerase Chain Reaction;
Viral Proteins;
immunology
- From:
Chinese Journal of Virology
2009;25(1):63-67
- CountryChina
- Language:Chinese
-
Abstract:
To construct Fv antibodies against H5N1 Avian influenza virus hemagglutinin,extracted mRNA from B lymphoblastoid cell lines secreting anti-HA antibodies was used and the VH and VL genes were amplified by RT-PCR and linked together by splicing overlap extension (SOE) with (Gly4 Ser)3 linker. The recombinant plasmid was then transformed to E. coli BL21(DE3) and sequence analysis indicated the total length of scFv was 714 bp and the expression of Fv was validated by PAGE and Western blot.
