[Cloning and expression of single-chain Fv antibodies against H5N1 Avian influenza virus hemagglutinin].
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.
Hao
Shu-mei
SM
Changchun University of Science and Technology, Changchun 130022, China. Haosm@sina.com
Li
Qun
Q
Wang
Cheng-yu
CY
Wang
Hua-lei
HL
Yang
Song-tao
ST
Xia
Xian-zhu
XZ
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D001717:Birds; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006388:Hemagglutinins / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism; D053124:Influenza A Virus, H5N1 Subtype / Q000187:drug effects / Q000276:immunology; D005585:Influenza in Birds / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014764:Viral Proteins / Q000276:immunology*
D000906:Antibodies; D006388:Hemagglutinins; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins