Loomis W F, Welker D, Hughes J, Maghakian D, Kuspa A
Department of Biology, University of California at San Diego, La Jolla 92093, USA.
Genetics. 1995 Sep;141(1):147-57. doi: 10.1093/genetics/141.1.147.
Detailed maps of the six chromosomes that carry the genes of Dictyostelium discoideum were constructed by correlating physically mapped regions with parasexually determined linkage groups. Chromosomally assigned regions were ordered and positioned by the pattern of altered fragment sizes seen in a set of restriction enzyme mediated integration-restriction fragment length polymorphism (REMI-RFLP) strains each harboring an inserted plasmid that carries sites recognized by NotI, SstI, SmaI, BglI and ApaI. These restriction enzymes were used to digest high molecular weight DNA prepared from more than 100 REMI-RFLP strains and the resulting fragments were separated and sized by pulsed-field gels. More than 150 gene probes were hybridized to blots of these gels and used to map the insertion sites relative to flanking restriction sites. In this way, we have been able to restriction map the 35 mb genome as well as determine the map position of more than 150 genes to with approximately 40 kb resolution. These maps provide a framework for subsequent refinement.
通过将物理图谱区域与准性生殖确定的连锁群相关联，构建了携带盘基网柄菌基因的6条染色体的详细图谱。通过在一组限制酶介导整合-限制片段长度多态性（REMI-RFLP）菌株中观察到的片段大小改变模式，对染色体分配区域进行排序和定位，每个菌株都含有一个插入质粒，该质粒携带被NotI、SstI、SmaI、BglI和ApaI识别的位点。这些限制酶用于消化从100多个REMI-RFLP菌株制备的高分子量DNA，所得片段通过脉冲场凝胶进行分离和大小测定。超过150个基因探针与这些凝胶的印迹杂交，并用于绘制相对于侧翼限制位点的插入位点图谱。通过这种方式，我们能够对35兆碱基的基因组进行限制酶图谱绘制，并以约40千碱基的分辨率确定150多个基因的图谱位置。这些图谱为后续的完善提供了框架。
