Sweeney G E, Watts D I, Turnock G
Nucleic Acids Res. 1987 Feb 11;15(3):933-45. doi: 10.1093/nar/15.3.933.
We have prepared cDNA libraries for amoebae and plasmodia of the acellular slime mould, Physarum polycephalum. Differential screening was used to isolate cell-type-specific cDNA clones (in bacteriophage M13) and both libraries yielded approximately 5% of such sequences. The amoebal- and plasmodial-specific clones were used to assay changes in transcription during the amoebal-plasmodial transition. The results obtained substantiate the view that the switch from amoebal to plasmodial characteristics occurs over several nuclear division cycles. With one exception, the specific cDNAs came from single-gene families. Southern blotting experiments also showed that they hybridised to identical restriction fragments from amoebal and plasmodial DNAs indicating that genomic rearrangements are unlikely to be involved in the regulation of these genes.
我们已经为多核变形虫(一种无细胞黏菌)的变形虫和原质团制备了互补DNA(cDNA)文库。采用差异筛选法分离细胞类型特异性的cDNA克隆(噬菌体M13中的克隆),两个文库中此类序列的比例均约为5%。利用变形虫特异性和原质团特异性克隆来检测变形虫-原质团转变过程中的转录变化。所得结果证实了以下观点:从变形虫特征向原质团特征的转变发生在几个核分裂周期中。除了一个例外,特异性cDNA均来自单基因家族。Southern印迹实验还表明,它们与来自变形虫和原质团DNA的相同限制性片段杂交,这表明基因组重排不太可能参与这些基因的调控。