Eggen A, Gautier M, Billaut A, Petit E, Hayes H, Laurent P, Urban C, Pfister-Genskow M, Eilertsen K, Bishop M D
Laboratoire de génétique biochimique et de cytogénétique, Département de génétique animale, Institut national de la recherche agronomique, 78352 Jouy-en-Josas Cedex, France.
Genet Sel Evol. 2001 Sep-Oct;33(5):543-8. doi: 10.1186/1297-9686-33-5-543.
A bovine artificial chromosome (BAC) library of 105 984 clones has been constructed in the vector pBeloBAC11 and organized in 3-dimension pools and high density membranes for screening by PCR and hybridization. The average insert size, determined after analysis of 388 clones, was estimated at 120 kb corresponding to a four genome coverage. Given the fact that a male was used to construct the library, the probability of finding any given autosomal and X or Y locus is respectively 0.98 and 0.86. The library was screened for 164 microsatellite markers and an average of 3.9 superpools was positive for each PCR system. None of the 50 or so BAC clones analysed by FISH was chimeric. This BAC library increases the international genome coverage for cattle to around 28 genome equivalents and extends the coverage of the ruminant genomes available at the Inra resource center to 15 genome equivalents.
已使用载体pBeloBAC11构建了一个包含105984个克隆的牛人工染色体(BAC)文库,并将其组织成三维池和高密度膜,用于通过聚合酶链反应(PCR)和杂交进行筛选。在对388个克隆进行分析后确定的平均插入片段大小估计为120 kb,相当于四个基因组覆盖度。鉴于构建文库使用的是雄性牛,找到任何给定常染色体以及X或Y基因座的概率分别为0.98和0.86。该文库针对164个微卫星标记进行了筛选,每个PCR系统平均有3.9个超级池呈阳性。通过荧光原位杂交(FISH)分析的约50个BAC克隆均无嵌合现象。这个BAC文库将牛的国际基因组覆盖度提高到约28个基因组当量,并将法国国家农业研究院(Inra)资源中心现有的反刍动物基因组覆盖度扩展到15个基因组当量。