Visser E S, Ambrosio R E, de Waal D T
Molecular Biology Section, Veterinary Research Institute, Onderstepoort, South Africa.
Vet Microbiol. 1991 Aug 15;28(3):313-25. doi: 10.1016/0378-1135(91)90086-u.
An Anaplasma centrale genomic library was constructed in pUC13. Two clones pAC5 and pAC137 hybridising to A. centrale and A. marginale DNA were isolated from this library. One of these, pAC5, also hybridised to DNA from A. ovis. The total insert of pAC5 was subcloned into pBR322. This subclone, pAC5-12, could detect 1 ng A. centrale, 0.5 ng A. marginale and 3.9 ng A. ovis DNA. The hybridisation pattern obtained with pAC5-12 on digests of A. centrale, A. marginale and A. ovis DNA suggests that this probe detects EcoR1 and Hind111-polymorphisms. Probe pAC5-12 could detect A. ovis DNA in 36% of blood samples tested compared to the 33% detectability obtained with microscopy.
构建了一个中央无形体基因组文库，载体为pUC13。从该文库中分离出两个与中央无形体和边缘无形体DNA杂交的克隆pAC5和pAC137。其中一个克隆pAC5还与绵羊无形体的DNA杂交。将pAC5的总插入片段亚克隆到pBR322中。这个亚克隆pAC5-12能够检测到1 ng中央无形体DNA、0.5 ng边缘无形体DNA和3.9 ng绵羊无形体DNA。用pAC5-12对中央无形体、边缘无形体和绵羊无形体DNA酶切产物进行杂交分析，结果表明该探针能检测到EcoR1和HindIII多态性。与显微镜检测33%的检出率相比，探针pAC5-12在36%的检测血样中能检测到绵羊无形体DNA。
