›› 2007, Vol. 50 ›› Issue (7): 655-661.

• RESEARCH PAPERS • Previous Articles     Next Articles

Expression and activity analysis of the recombinant Solenopsis invicta Buren venom allergen Sol i1

HAN Xue-Qing, LIN Xiang-Mei, ZHANG Yong-Guo, CHEN Yan, YE Gui-Sheng, YANG Wei-Dong,XIA Qiao-Yu, YANG Ze-Xiao, WANG Jian-Feng   

  1. (Chinese Academy of Inspection and Quarantine, Beijing 100029, China)
  • Online:2007-08-21 Published:2007-08-21
  • Contact: HAN Xue-Qing

Abstract: ObjectiveTo study the molecular pathogenic mechanism of Solenopsis invicta Buren venom allergens and produce reagents for the prophylaxis and therapy of S. invicta sting. Methods In the study, the S. invicta venom allergens sol i1 gene and its fragment soli1a encoding the truncated peptide without transmembrane region were amplified by RT-PCR and nPCRand then be sequenced. Two recombinant plasmids Sol i1-pET43.1a and Sol i1a-pET43.1a were constructed and verified by PCR, RE digestion and sequencing, and then transformed into E. coli BL21(DE3) and induced by IPTG. The expression products were purified by affinity chromatograph following with the identification of SDS-PAGE and Western blotting, and then used to inoculate rabbits for analyzing the allergenic activity and the potential in therapy. Results The results showed that the nucleotide homology of the amplified product with that published in GenBank was 99%, and the recombinant fusion proteins expressed in BL21(DE3) at high-level had good allergenic activity and immunotherapy potential. Conclusions Recombinant Sol i1 (rSol i1) and recombinant Sol i1a (rSol i1a) expressed in BL21(DE3) both had excellent biological activity, which set basis for further studying the mechanism of S. invicta venom allergens and the prevention of S. invicta.

Key words: Solenopsis invicta, gene, venom allergens, expression, biological activity analysis