ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2005, Vol. 36 ›› Issue (7): 705-710.doi:

• 兽医 • Previous Articles     Next Articles

Immunogenic Characterization of the Recombinant Attenuated Salmonellatyphimurium Expressing MIC2 Gene of Eimeria necatrix

XIE Ming-quan; QIN Zong-hua;CAI Jian-ping;Ahmed Bashar Elhag;
PENG Xin-yu;WEI Wen-kang;WU Hui-xian   

  1. 1.Institute of Veterinary Medicine, Guangdong Academy of Agricultral Sciences, Guangzhou 510640,China; 2. School of Veterinary Science, Nyala University, Sudan
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-07-25 Published:2005-07-25

Abstract: The ORF sequence of micronem protein-2(MIC2) gene from Eimeria necatrix(GD) was cloned by PCR amplification using specific primers designed according to the sequence of EnMIC2(GD), and ligated to vector pYA3342 to construct recombinant plasmid pYA3342-EnMIC2 for transforming E.coli x6212. Then the positive recombinant plasmid identified by PCR amplification and endonuclease digestion was electroporated into attenuated S. typhimurium X4550. The recombinant EnMIC2 expressed in Salmonella by inducing of 1 mmol/L IPTG was approximately 35 ku in size and accounting for 8.6% in total bacterial proteins and could be detected in western blotting using hyperimmune serum against E.necatrix. The specific serum IgG against EnMIC2 in chickens post-immunized(PI) with recombinant Salmonella at the dosage of 108 or 109 CFU per bird was probed, and reached to the peak 3 weeks PI. Specific IgA in intestine was also detected 3 weeks PI. These immunized chicken were challenged with 500 E. necatrix sporulated oocysts per bird 3 weeks PI and can reduce the oocyst production by 26.62% and 48.92%, respectively in low or high vaccinating dosage compared to the control groups.