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| Preparation of Two Different Lengths of FopA Antigen and Antibody Used in Francisella tularemia Detecting |
| JING Ying-ying1,2, YANG Yu1, WANG Jing1, YANG Yong-li1, HU Kong-xin1, WANG Zhen-dong2 |
1. Chinese Academy of Inspection and Quarantine, Beijing 100123, China;
2. College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China |
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Abstract Objective:To establish an BL21 expression system of Francisella tularemia-specific antigen (FopA-L and FopA-S), then obtain their highly active recombinant protein and the polyclonal antibodies which can be applied for Francisella monitoring, diagnosis and treatment.Methods:Constructing a plasmid expression vector pET100 FopA-L and FopA-S, then transforming FopA plasmid into E. coli BL21 cells, induced by IPTG, purifing protein by chelate nickel ion-NTA (Ni-NTA) affinity chromatography, making polyclonal antibody by recombinant FopA-L and FopA-S proteins immune rabbits and antibody were determined by Western blot, ELISA and GICA. Results:Constructed expression vectors pET100 of FopA-L and FopA-S, the BL21 cell lines obtained can highly express target proteins, antibody specifically combined with them which can be used in Francisella detecting were prepared successfully, with a titer of more than 1 ∶100000 and highly specific. Conclusion:Preparation the FopA-L and FopA-S antigen and antibody laid the foundation for the establishment of rapid detection method of Francisella.
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Received: 06 August 2010
Published: 25 December 2010
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