Piñol J, Cabré O
Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Gene. 1994 May 27;143(1):39-41. doi: 10.1016/0378-1119(94)90601-7.
In order to take advantage of non-radioactive methods, we have developed two plasmids (p lambda LE and p lambda RE) for mapping restriction sites of long inserts cloned in phage lambda vectors. These plasmids are constructed by cloning the left 402-bp and right 560-bp phage lambda genome ends, respectively. To map restriction sites, the cloned sequences in p lambda LE and p lambda RE are labeled with digoxygenin and hybridized to partially digested lambda DNA. The ladder of bands detected with these probes can be used to construct restriction maps in the same way as those obtained using radioactively labeled cos complementary oligodeoxyribonucleotides [Rackwitz et al., Gene 30 (1984) 195-200].
为了利用非放射性方法，我们开发了两种质粒（pλLE和pλRE），用于绘制克隆在噬菌体λ载体中的长插入片段的限制性酶切位点图谱。这些质粒分别通过克隆噬菌体λ基因组左侧402 bp和右侧560 bp的末端构建而成。为了绘制限制性酶切位点图谱，将pλLE和pλRE中的克隆序列用地高辛标记，并与部分消化的λDNA杂交。用这些探针检测到的条带阶梯可用于构建限制性图谱，其方式与使用放射性标记的粘性末端互补寡脱氧核糖核苷酸获得的图谱相同[拉克维茨等人，《基因》30（1984）195 - 200]。
