Piper M B, Bankier A T, Dear P H
Medical Research Council Laboratory of Molecular Biology, Protein and Nucleic Acid Chemistry Division, Cambridge CB2 2QH, UK.
Genome Res. 1998 Dec;8(12):1299-307. doi: 10.1101/gr.8.12.1299.
We have constructed a HAPPY map of the apicomplexan parasite Cryptosporidium parvum. We have placed 204 markers on the 10.4-Mb genome, giving an average marker spacing of approximately 50 kb, with an effective resolution of approximately 40 kb. HAPPY mapping (an in vitro linkage technique based on screening approximately haploid amounts of DNA by the polymerase chain reaction) is fast and accurate and is not subject to the distortions inherent in cloning, meiotic recombination, or hybrid cell formation. In addition, little genomic DNA is needed as a substrate, and the AT content of the genome is largely immaterial, making it an ideal method for mapping otherwise intractable parasite genomes. The map, covering all eight chromosomes, consists of 10 linkage groups, each of which has been chromosomally assigned. We have verified the accuracy of the map by several methods, including the construction of a >140-kb PAC contig on chromosome VI. Less than 1% of our markers detect non-rDNA duplicated sequences.
我们构建了顶复门寄生虫微小隐孢子虫的HAPPY图谱。我们在10.4兆碱基的基因组上放置了204个标记，平均标记间距约为50千碱基，有效分辨率约为40千碱基。HAPPY图谱构建（一种基于通过聚合酶链反应筛选大约单倍体量DNA的体外连锁技术）快速且准确，不受克隆、减数分裂重组或杂交细胞形成中固有的扭曲影响。此外，作为底物所需的基因组DNA很少，并且基因组的AT含量在很大程度上无关紧要，这使其成为绘制其他难以处理的寄生虫基因组图谱的理想方法。该图谱覆盖了所有八条染色体，由10个连锁群组成，每个连锁群都已进行了染色体定位。我们通过多种方法验证了图谱的准确性，包括在第六条染色体上构建一个大于140千碱基的PAC重叠群。我们的标记中不到1%检测到非核糖体DNA重复序列。
