Tripp C A, Wagner G G, Rice-Ficht A C
Department of Veterinary Microbiology and Parasitology, Texas A&M University, College Station 77843.
Exp Parasitol. 1989 Oct;69(3):211-25. doi: 10.1016/0014-4894(89)90068-4.
Genomic DNA prepared from erythrocyte cultures of Babesia bovis merozoites was digested with mung bean nuclease and used to construct a lambda gt11 expression library of B. bovis recombinants. Immunoscreening with two polyclonal antibody probes detected multiple recombinants from which two, designated Bb-1 and Bb-3, were chosen for further analysis. Monospecific immunoglobulins isolated from the screening sera using nitrocellulose-bound fusion proteins were employed to determine the native molecular weight and the intracellular location of the babesial proteins encoded by the recombinants. Clone Bb-1 encodes an antigen of 77,000 Da located at the apical end of the intraerythrocytic parasite. A protein of 75,000 Da encoded by clone Bb-3 is associated with the infected red blood cell cytoplasm and/or membrane but not with the merozoite.
从牛巴贝斯虫裂殖子的红细胞培养物中制备的基因组DNA用绿豆核酸酶消化，并用于构建牛巴贝斯虫重组体的λgt11表达文库。用两种多克隆抗体探针进行免疫筛选，检测到多个重组体，从中选择了两个命名为Bb-1和Bb-3的重组体进行进一步分析。使用与硝酸纤维素结合的融合蛋白从筛选血清中分离出的单特异性免疫球蛋白，用于确定重组体编码的巴贝斯虫蛋白的天然分子量和细胞内定位。克隆Bb-1编码一种77,000道尔顿的抗原，位于红细胞内寄生虫的顶端。克隆Bb-3编码的一种75,000道尔顿的蛋白质与感染的红细胞细胞质和/或膜相关，但与裂殖子无关。
