Kathir Pushpa, LaVoie Matthew, Brazelton William J, Haas Nancy A, Lefebvre Paul A, Silflow Carolyn D
Department of Genetics, University of Minnesota, St. Paul, Minnesota, USA.
Eukaryot Cell. 2003 Apr;2(2):362-79. doi: 10.1128/EC.2.2.362-379.2003.
We have prepared a molecular map of the Chlamydomonas reinhardtii genome anchored to the genetic map. The map consists of 264 markers, including sequence-tagged sites (STS), scored by use of PCR and agarose gel electrophoresis, and restriction fragment length polymorphism markers, scored by use of Southern blot hybridization. All molecular markers tested map to one of the 17 known linkage groups of C. reinhardtii. The map covers approximately 1,000 centimorgans (cM). Any position on the C. reinhardtii genetic map is, on average, within 2 cM of a mapped molecular marker. This molecular map, in combination with the ongoing mapping of bacterial artificial chromosome (BAC) clones and the forthcoming sequence of the C. reinhardtii nuclear genome, should greatly facilitate isolation of genes of interest by using positional cloning methods. In addition, the presence of easily assayed STS markers on each arm of each linkage group should be very useful in mapping new mutations in preparation for positional cloning.
我们已经制备了一张与遗传图谱锚定的莱茵衣藻基因组分子图谱。该图谱由264个标记组成,包括通过PCR和琼脂糖凝胶电泳进行评分的序列标签位点(STS),以及通过Southern杂交进行评分的限制性片段长度多态性标记。所有测试的分子标记都定位到莱茵衣藻17个已知连锁群中的一个。该图谱覆盖约1000厘摩(cM)。莱茵衣藻遗传图谱上的任何位置平均距离已定位的分子标记在2 cM以内。这张分子图谱,结合正在进行的细菌人工染色体(BAC)克隆图谱绘制以及即将公布的莱茵衣藻核基因组序列,应该会极大地促进通过定位克隆方法分离感兴趣的基因。此外,每个连锁群每条臂上都存在易于检测的STS标记,这对于为定位克隆准备而进行新突变的图谱绘制将非常有用。