Rouquier S, Batzer M A, Giorgi D
Human Genome Center, Lawrence Livermore National Laboratory, Livermore, California 94551.
Anal Biochem. 1994 Mar;217(2):205-9. doi: 10.1006/abio.1994.1110.
In order to increase the rate of generation of a contiguous human chromosome 19 physical map, we have investigated the advantages of using a bacterial artificial chromosome (BAC) library in comparison with other systems. This cloning system, recently described, can faithfully propagate DNA fragments greater than 300 kb in size. We have screened a total human genomic BAC library with a complex cosmid probe, specific for the ryanodine receptor gene (RYR1) located at 19q13.1. One 150-kb BAC was positive for the ryanodine receptor probe. The ryanodine receptor BAC hybridized to a 150-kb overlapping set of nine chromosome 19-specific cosmids at the 3' terminus of the RYR1 gene as well as two established cosmid contigs which can be linked to the same physical region. The hybridization of the BAC to a 150-kb set of chromosome 19 cosmids suggests that this BAC is nonchimeric in structure.
为了提高连续人类19号染色体物理图谱的构建速度，我们研究了使用细菌人工染色体（BAC）文库相较于其他系统的优势。这种克隆系统最近被报道，能够忠实地扩增大小超过300 kb的DNA片段。我们用一个复杂的黏粒探针筛选了一个全人类基因组BAC文库，该探针针对位于19q13.1的兰尼碱受体基因（RYR1）。一个150 kb的BAC对兰尼碱受体探针呈阳性反应。该兰尼碱受体BAC与位于RYR1基因3'端的一组九个19号染色体特异性黏粒的150 kb重叠片段杂交，以及与两个可连接到同一物理区域的已建立的黏粒重叠群杂交。BAC与一组150 kb的19号染色体黏粒的杂交表明该BAC在结构上是非嵌合的。
