Integrated maps of the chromosomes in Dictyostelium discoideum.
Integrated maps of the chromosomes in Dictyostelium discoideum.
复制标题
盘基网柄菌染色体的整合图谱。
DOI：
10.1093/genetics/141.1.147
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发表时间：
1995
期刊：
影响因子：
3.3
通讯作者：
Kuspa,A
中科院分区：
文献类型：
--
作者：
Loomis,WF;Welker,D;Hughes,J;Maghakian,D;Kuspa,A
通过将物理映射的区域与准性决定的连锁群相关联，构建了携带盘状盘齿龙基因的6条染色体的详细图谱。在一组限制性内切酶介导的整合-限制性片段长度多态性（REMI-RFLP）菌株中，通过改变片段大小的模式对染色体分配区域进行排序和定位，每个菌株都含有插入的质粒，该质粒携带NotI， SstI, SmaI， BglI和ApaI识别的位点。这些限制性内切酶用于消化从100多个REMI-RFLP菌株中制备的高分子量DNA，并通过脉冲场凝胶分离所得片段并确定大小。将150多个基因探针杂交到这些凝胶的印迹上，并用于绘制相对于侧翼限制性内切位点的插入位点。通过这种方法，我们已经能够以大约40kb的分辨率绘制35mb基因组的限制性图谱，并确定150多个基因的图谱位置。这些映射为后续的细化提供了一个框架。
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.