Evaluation of the immunoprotective effect of E.tenella recombinant Profilin protein in chickens
[Objective]In this study,we investigated the immunoprotective effect of recombinant Profilin protein on chickens by prokaryotic expression and purification of E.tenella Profilin gene.[Methods]The Profilin gene specific primers were designed and synthesized,the total RNA of E.enella spore oocyst was extracted and reverse-transcribed into cDNA.Profilin gene was obtained by PCR amplification and then connected to pET-32a(+)vector to construct the recombinant plasmid pET-32a(+)-Profilin.It was expressed and purified under IPTG induction.Survival rate,mean weight gain,blood stool count,number of oo-cysts per gram of faeces(OPG),oocysts production,cecal lesions,and anti-coccidia index(ACI)were measured and calculated for chickens immunized with recombinant Profilin protein,followed by oral administration of Eimeria tender oocysts.The his-topathological changes of cecum were observed and the levels of IL-2,IL-10,TNF-α and sIgA in serum were determined to eval-uate the immunoprotective effect of recombinant Profilin protein.[Results]Recombinant Profilin protein with a size of 510bp was successfully expressed.Immune protection test showed that the survival rate,blood stool count and OPG of chickens in Profilin recombinant protein group were significantly decreased compared with those in non-immune attack group(P＜0.05).The 50 μg recombinant protein immunogroup had the best body weight gain at 93％,the least oocyst production,and the best ACI results.The 100 μg recombinant protein-immunized group showed the least damage to the cecum,whereas no significant difference was observed in the immune organ index(P＞0.05).At7days after attack,serum levels of IL-2,TNF-a,IL-10 and sIgA in the non-immune attack group were significantly higher than those in the blank control group(P＜0.01).The levels of IL-2,TNF-a,IL-10 and sIg A in the recombinant protein immune group were significantly lower than those in the non-immune group(P＜0.01).[Conclusion]Recombinant Profilin protein can induce cellular immunity and humoral immunity in the body,and shows good anti-coccidial effect in E.tenella infection.
万方数据
维普
