文章信息：Li Yi and Shipeng Cheng
Institute of Special Wild Economic Animal and Plant Science, Chinese Academy of Agricultural Sciences, Changchun, Jilin Province, P.R. China.
Address correspondence to: Shipeng Cheng Institute of Special Wild Economic Animal and Plant Science Chinese Academy of Agricultural Sciences4899 Juye Street Changchun Jilin Province
P.R.China 130122 E-mail:tcscsp@126.com
Received: January 26, 2015 Accepted: April 9, 2015
原文摘要：
The variable regions of the heavy chain (VH) and light chain (VL) were amplified by RT-PCR from the hybridoma 1N8, which secretes the monoclonal antibody against CDV N protein (aa 277-471). The VL and VH amplicons were combined using SOE-PCR by a 12 amino acid flexible linker (SSGGGGSGGGGS), which produced the scFv gene (named scFv/1N8). After sequence analysis, the scFv/1N8 gene was cloned into the prokaryotic expression vector PET32a with a His-tag. The recombinant scFv/1N8 protein was successfully expressed in recombinant Escherichia coli by IPTG induction. Moreover, the binding activity and specificity of the scFv were determined by indirect ELISA (His-tag) and competitive ELISA. The recombinant scFv/1N8 protein reported here will provide some basis for further antiviral drug research based on the scFv molecule.