Jimenez-Ruiz A, Requena J M, Lancillotti F, Morales G, Lopez M C, Alonso C
Centro de Biologia Molecular, CSIC-UAM, Madrid, Spain.
Nucleic Acids Res. 1989 Jun 26;17(12):4579-88. doi: 10.1093/nar/17.12.4579.
We describe the selection of a group of plasmids with potential to form Z-DNA, from libraries of Drosophila hydei nuclear DNA using anti Z-DNA monoclonal (22) or polyclonal (10c) antibodies. The supercoiled closed circular forms of most of the selected recombinant plasmids from the 10c Z-DNA library show affinity to the polyclonal 10c antibody as indicated by DNA binding assays. One of these plasmids, pF17, was selected for further study. The insert in this plasmid adopts the Z conformation at bacterial supercoiled density. Analysis of deletion plasmids indicates that a Z-epitope is located within a short fragment of the insert in which 3 GC repetitions are found. The Drosophila DNA insert in pF17 hybridizes in situ with locus 4-75C1-2 of the polytene chromosomes, a locus whose transcriptional activity is developmentally regulated during the third instar.
我们描述了使用抗Z-DNA单克隆抗体（22）或多克隆抗体（10c），从海德果蝇核DNA文库中筛选出一组具有形成Z-DNA潜力的质粒的过程。如DNA结合分析所示，来自10c Z-DNA文库的大多数选定重组质粒的超螺旋闭环形式对多克隆10c抗体具有亲和力。其中一个质粒pF17被选作进一步研究。该质粒中的插入片段在细菌超螺旋密度下采用Z构象。缺失质粒分析表明，一个Z表位位于插入片段的一个短片段内，该片段中有3个GC重复序列。pF17中的果蝇DNA插入片段与多线染色体的4-75C1-2位点原位杂交，该位点的转录活性在三龄幼虫期受到发育调控。
