CJCU

Chem. J. Chinese Universities ›› 2011, Vol. 32 ›› Issue (1): 84.

• Articles • Previous Articles     Next Articles

Expression and Preliminary Activity Identification of Several Linked Antigen Epitopes of Consensce Genes of Yersinia Pestis, Hantavirus and Leptospira

PU Yun1,2, QI Yan-Fei1, XU Guo-Liang3, XU Hui3, ZHANG Shi-Yao1, MENG Ri-Zeng1,2, SONG Xiu-Ling1, YANG Huai-Ning2, LI Juan1*   

  1. 1. School of Public Health, Jilin University, Changchun 130021, China;
    2. Jilin Entry & Exit Inspection and Quarantine Bureau, Changchun  130062,   China;
    3.  Cardiovascular Center, First Hospital of Jilin University, Changchun 130021, China
  • Received:2010-06-13 Revised:2010-10-08 Online:2011-01-10 Published:2010-12-11
  • Contact: LI Juan E-mail:Li_juan@jlu.edu.cn
  • Supported by:

    国家“重大新药创制”科技重大专项项目(批准号: 2009ZX09103-105)、高等学校博士学科点专项科研基金(批准号: 20090061120093)、国家质量监督检验检疫总局科技计划项目(批准号:  2009IK214)和吉林大学研究生创新研究计划项目(批准号:  20091028)资助.

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Abstract: The expression vector pET-rYHL was constructed by inserting the linked gene contained two antigen epitopes of F1 capsule antigen from Yersinia Pestis, six antigen epitopes of Hantavirus and six antigen epitopes of Leptospira into pET-20b and was identified by digestion with restriction enzymes and sequence analysis. Then an expression strain was selected after transformation of the recombined plasmid into E. coli BL21 (DE3), fusion protein with His-tag was efficiently expressed in the form of inclusion body after IPTG induction. The inclusion body was washed, dissolved and purified by Ni2+ chelate chromatography under denatured condition. SDS-PAGE analysis and Western blotting showed that the fusion protein with a molecular weight of about 40 000 was purified and the purity was up to 98%. The results ELISA showed specific reactions with Plague, Epidemic hemorrhagic fever and Leptospirosis positive sera respectively, and no cross-reaction with other positive sera sample(salmonella ) using the expression protein.

Key words: Plague, Hantavirus, Leptospira, Chimeric gene, Gene expression, Activity identification

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