Tetzlaff C L, McMurray D N, Rice-Ficht A C
Department of Medical Microbiology and Immunology, Texas A&M University, College Station 77843.
Mol Biochem Parasitol. 1990 May;40(2):183-92. doi: 10.1016/0166-6851(90)90040-s.
Babesia microti genomic DNA was purified from parasitized murine erythrocytes, digested with mung bean nuclease and used to construct an expression library in lambda gt11. Polyspecific antisera from mice infected with virulent B. microti organisms (ATCC30221) were used to screen the genomic library for genes encoding major immunogens. High titer antisera selected a recombinant phage, Bm13, containing 3.3 kb of B. microti DNA. Hybridization analysis confirmed the parasite origin of the clone; affinity-purified antibody revealed a native molecular weight of 54,000 for the B. microti protein encoded by the recombinant. Only genomic DNA isolated from the virulent strain of B. microti contained sequences which hybridized to Bm13. Genomic DNA prepared from the Peabody attenuated strain of B. microti or from Babesia bovis DNA did not contain any complementary sequences. These data suggest a possible role for the gene in the virulence of the organism.
微小巴贝斯虫基因组DNA从感染的鼠红细胞中纯化出来，用绿豆核酸酶消化，用于构建λgt11表达文库。用感染强毒微小巴贝斯虫（ATCC30221）的小鼠产生的多特异性抗血清筛选基因组文库，以寻找编码主要免疫原的基因。高滴度抗血清筛选出一个重组噬菌体Bm13，它含有3.3 kb的微小巴贝斯虫DNA。杂交分析证实了该克隆来源于寄生虫；亲和纯化抗体显示重组体编码的微小巴贝斯虫蛋白天然分子量为54,000。只有从微小巴贝斯虫强毒株分离的基因组DNA含有与Bm13杂交的序列。从皮博迪减毒株微小巴贝斯虫或牛巴贝斯虫DNA制备的基因组DNA不含有任何互补序列。这些数据表明该基因可能在该生物体的毒力中起作用。
