摘要
本研究初步评价了柔嫩艾美耳球虫热休克蛋白70(EtHSP70)的免疫保护效果。以RT-PCR方法克隆获得E.tenella广东株的Ethsp70基因序列,插入表达载体pMAL-c2X后转化入大肠杆菌Rosetta株,经IPTG诱导,可高效表达分子量为112 ku的可溶性融合蛋白,表达量约占菌体总蛋白的27%。将Ethsp70基因插入真核载体pcDNA6,构建真核表达质粒pcDNA-Ethsp70。用纯化的重组融合蛋白(rEtHSP70)和pcDNA6-Ethsp70质粒分别以100μg/只剂量肌注免疫雏鸡,攻虫后以盲肠病变计分、相对盲肠卵囊产量(ROP)、相对增重率和抗球虫指数(ACI)为评价指标,结果表明2个免疫组相对盲肠卵囊产量分别为39.4%、45.2%,抗球虫指数由89分别提高至免疫组的164和150。提示EtHSP70可能是一种有潜在应用价值的保护性抗原。
The immuno-protective effects of heat shock protein 70 from Eimeria tenella （EtHSP70） was evaluated in this study. The EtHSP70 gene fragment was obtained by RT-PCR with a pair of specific primers designed according to Ethsp70 gene sequence in GenBank, and inserted into the plasmid pMAL-c2X and pcDNA6 for constructing recombinant expressing vector pMAL-c2X-Ethsp70 and pcDNA6-Ethsp70, respectively. Recombinant pMAL-c2X-Ethsp70 was induced with IPTG to express the fusion EtHSP70 in the transformed E. coli Rosseta strain. Recombinant EtHSP70 with a molecular weight of 112 ku accounted for 27% of the total bacterial proteins. Every chicken in different groups was separately vaccinated with 100 Ixg of recombinant fusion EtHSP70 or pcDNA6-Ethsp70 at 14 and 21 day-old, and challenged with sporolated occysts of E. tenella at 28 day-old. Some indexes, including relative weight gain, relative oocyst production （ROP） in caeca, and anticoccidial index （ACI） , were used to evaluate the immune effect. The results showed that the ROP is 39.4% and 45.2% in rEtHSP70 vaccinated group and pcDNA6-Ethsp70 immunized group, respectively. It suggested that partial protection against the challenge of E. tenalla oocysts could be triggered after the chicken was vaccinated with the vaccines based on EtHSP70.
出处
《畜牧与兽医》
北大核心
2008年第10期31-36,共6页
Animal Husbandry & Veterinary Medicine
基金
国家高科技发展计划项目(“863”)资助,项目编号:2006AA10A207
