CAJ | 학술논문

为评价gain56基因重组表达产物作为亚单位疫苗预防鸡巨型艾美球虫(E.maxima),感染的效果,以E.maxima NT株配子体总RNA为模板,RT.PCR扩增和克隆gain56基因,选择该基因两段丰富抗原表位的编码区片段.利用原核表达载体pGEX-6P-1在大肠杆菌中进行截短表达.以可溶性的GST-gain56-2融合蛋白为免疫原,设立高、中、低(1.0 mg、0.5 mg、0.25 mg)3个剂量,单独或使用弗氏完全佐剂,于5 d、12 d和19 d对雏鸡进行3次免疫,26 d用5 ×10~4个E.maxima孢子化卵囊进行攻虫,8 d后迫杀,对各组的存活率、相对增重率、卵囊减少率、ACI等指标进行统计分析.结果显示:就相对增重率而言各免疫组比未免疫攻毒组的高27.6%以上,而各免疫组之间差异不显著(p>0.05);就卵囊减少率指标而言,各免疫组较未免疫攻毒组均有不同程度减少,但是均小于75%;就ACI指标而言,各免疫组较未免疫攻毒组均有不同程度增加,以中剂量蛋白佐剂组为最高.GST-gam56-2融合蛋白对于E.maxima感染具有一定的免疫保护效力.
위평개gain56기인중조표체산물작위아단위역묘예방계거형애미구충(E.maxima),감염적효과,이E.maxima NT주배자체총RNA위모판,RT.PCR확증화극륭gain56기인,선택해기인량단봉부항원표위적편마구편단.이용원핵표체재체pGEX-6P-1재대장간균중진행절단표체.이가용성적GST-gain56-2융합단백위면역원,설립고、중、저(1.0 mg、0.5 mg、0.25 mg)3개제량,단독혹사용불씨완전좌제,우5 d、12 d화19 d대추계진행3차면역,26 d용5 ×10~4개E.maxima포자화란낭진행공충,8 d후박살,대각조적존활솔、상대증중솔、란낭감소솔、ACI등지표진행통계분석.결과현시:취상대증중솔이언각면역조비미면역공독조적고27.6%이상,이각면역조지간차이불현저(p>0.05);취란낭감소솔지표이언,각면역조교미면역공독조균유불동정도감소,단시균소우75%;취ACI지표이언,각면역조교미면역공독조균유불동정도증가,이중제량단백좌제조위최고.GST-gam56-2융합단백대우E.maxima감염구유일정적면역보호효력.
To assess the protective effect of recombinant gain56 against E.maxima infection in chickens, gara56 gene of E.maxima was amplified by RT-PCR from total RNA of purified gametocytes. Two fragments with rich antigenic epitopes were subcloned into pGEX-6P-1 and expressed. Five-day old chickens were subcutaneously immunized with soluble GST-gam56-2 antigen with or without adjuvant, and boosted with the same dosage at 12, 19 days of age. All chickens were orally challenged with homologous sporulated oocysts at 26 days of age and killed at eight days post challenge. Weight gains, anticoccidial index and oocyst output were determined and compared with those of the control animals. The results showed the weight gains of the immunized chickens were significantly higher than that of the control group (p<0.05). The oocyst outputs in immunized chickens dropped by 35.92 to 57.75 percent compared with the controls. It was concluded that recombinant protein GST-gam56-2 could induce partial protection against E.maxima infection.