Miller G A, Bhogal B S, McCandliss R, Strausberg R L, Jessee E J, Anderson A C, Fuchs C K, Nagle J, Likel M H, Strasser J M
Molecular Biology Department, A. H. Robins Co., Richmond, Virginia 23220.
Infect Immun. 1989 Jul;57(7):2014-20. doi: 10.1128/iai.57.7.2014-2020.1989.
A cDNA clone derived from sporulated oocysts of Eimeria tenella and encoding the expression product GX3262 was identified using a monoclonal antibody raised against Eimeria acervulina sporozoites. The cDNA fragment containing the coccidial antigen gene was cloned in bacteriophage lambda gt11, transferred to a plasmid, and introduced into Escherichia coli for analysis of the gene products. The strain carrying the plasmid produced GX3262 as part of a fusion protein consisting of the first 1,006 amino acids of E. coli beta-galactosidase and 112 amino acids of the E. tenella protein of approximately 12 kilodaltons. Partially purified antigen, heat-killed recombinant bacterin, and live E. coli containing the recombinant coccidial antigen were used to immunize 1-week-old or newly hatched broiler chicks. Several immunization protocols were utilized, including boosts with partially purified beta-galactosidase-GX3262, bacterin, or small numbers of live E. tenella oocysts. After challenge with an experimental E. tenella infection, the birds were evaluated by scoring cecal lesions to determine the level of protection. The greatest degree of protection was seen after only a single immunization of 2-day-old birds with a live recombinant E. coli preparation. The results presented here identify GX3262 as a potential candidate coccidial vaccine antigen and provide evidence for the first time that newly hatched chickens can be successfully vaccinated with a recombinant antigen.
利用针对堆型艾美耳球虫子孢子产生的单克隆抗体,鉴定出一个源自柔嫩艾美耳球虫孢子化卵囊的cDNA克隆,该克隆编码表达产物GX3262。将含有球虫抗原基因的cDNA片段克隆到噬菌体λgt11中,转移到质粒上,然后导入大肠杆菌以分析基因产物。携带该质粒的菌株产生了GX3262,它是一种融合蛋白的一部分,该融合蛋白由大肠杆菌β-半乳糖苷酶的前1006个氨基酸和大约12千道尔顿的柔嫩艾美耳球虫蛋白的112个氨基酸组成。部分纯化的抗原、热灭活的重组菌苗和含有重组球虫抗原的活大肠杆菌被用于免疫1周龄或刚孵出的肉仔鸡。采用了几种免疫方案,包括用部分纯化的β-半乳糖苷酶-GX3262、菌苗或少量活的柔嫩艾美耳球虫卵囊进行加强免疫。在用实验性柔嫩艾美耳球虫感染进行攻毒后,通过对盲肠病变进行评分来评估鸡群,以确定保护水平。仅用活的重组大肠杆菌制剂对2日龄鸡进行一次免疫后,就观察到了最大程度的保护。本文的结果确定GX3262为一种潜在的球虫疫苗候选抗原,并首次提供了新孵出的鸡可用重组抗原成功免疫的证据。