PDF(651 KB)
Preparation of Two Different Lengths of FopA Antigen and Antibody Used in Francisella tularemia Detecting
JING Ying-ying, YANG Yu, WANG Jing, YANG Yong-li, HU Kong-xin, WANG Zhen-dong
China Biotechnology ›› 2010, Vol. 30 ›› Issue (12) : 76-81.
PDF(651 KB)
PDF(651 KB)
Preparation of Two Different Lengths of FopA Antigen and Antibody Used in Francisella tularemia Detecting
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.
Francisella tularemia / FopA / E.coli / Expression / Polyclonal antibody {{custom_keyword}} /
[1] McLendon M K, Apicella M A, Allen L A. Francisella tularensis: taxonomy, genetics, and Immunopathogenesis of a potential agent of biowarfare. Annu Rev Microbiol, 2006, 60: 167-185.
[2] Hopla C E. The ecology of tularemia.Adv Vet Sci Comp Med, 1974, 18: 25-53.
[3] 宫英, 逄增昌, 强新,等. 土拉弗氏菌病患者血清抗体20年变化趋势分析. 中国公共卫生, 2006, 22(10): 1201-1202. Gong Y, Pang Z C, Qiang X, et al. Chinese Journal of Public Health, 2006, 22(10): 1201-1202.
[4] Porsch-Ozcurumez M, Kischel N, Priebe H, et al. Comparison of enzyme-linked immunosorbent assay, Western blotting, microagglutination, indirect immunofluorescence assay, and flow cytometry for serological diagnosis of tularaemia. Clinical and Diagnostic, 2004, 11(6): 1001-1015.
[5] Aronova N V, Pavlovich N V. The use if lipopolysaccharide in the dot solid phase enzyme immunoassay. Mikrobiol Epidemiol Immunobiol, 2000, 5: 75-78.
[6] Berdal B P, Mehl R, Haaheim H, et al. Field detection of Francisella tularensis.Scand J Infect Dis, 2000, 32(3): 287-291.
[7] Magnarelli L, Levy S, Koski R. Detetion of antibodies to Francisella tularensis in cats.Rvsc, 2007, 82(1): 22-26.
[8] Tarnvik A, Berglund L. Tularemia. Eur Respir J, 2003, 21(2): 361-373.
[9] 靖学芳, 安云庆. LPS和抗LPS治疗的研究及应用进展. 微生物学免疫学进展, 2004, 32(2): 53-57. Jing X F, An Y Q. Progress in Microbiology and Immunology, 2004, 32(2): 53-57.
[10] Huntley J F, Conley P G, Hagman K E, et al. Characterization of Francisella tularensis outer membrane proteins. J Bacteriol, 2007, 189(2): 561-574.
[11] Fulop M, Manchee R, Titball R. Role of lipopolysaccharide and a major outer membrane protein from Francisella tularensis in the induction of immunity against tularemia. Vaccine, 1995, 13(13):1220-1225.
[12] Saier M H, Werner P K, Muller M. Insertion of proteins into bacterial membranes: mechanis, characteristics, and comparisons with the eucaryotic process. Microbiol Rev, 1989, 53(3): 333-366.
[13] Gunnar von Heijne. Life and death of a signal peptide. Nature, 1998, 396 (12):111-113.
[14] 何斌, 端青. 重组土拉弗朗西斯菌外膜蛋白 FopA 的融合表达及抗原性分析. 生物技术通讯, 2010, 21(1): 32-34. He B, Duan Q. Letters in Biotechnology, 2010, 21(1): 32-34.
[15] 王增, 马会勤, 张文,等. 包涵体蛋白的分离和色谱法体外复性纯化研究进展. 中国生物工程杂志, 2009, 29(7): 102-107. Wang Z, Ma H Q, Zhang W, et al.China Biotechnology, 2009, 29(7): 102-107.
[16] 张婷婷, 叶波平. 包涵体蛋白质的复性研究进展. 药物生物技术, 2007, 14 (4): 306-309. Zhang T T, Ye B P. Pharmaceutical Biotechnology, 2007, 14 (4): 306-309.
PDF(651 KB)
/
| 〈 |
|
〉 |
