Willmitzer L, Otten L, Simons G, Schmalenbach W, Schröder J, Schröder G, Van Montagu M, de Vos G, Schell J
Mol Gen Genet. 1981;182(2):255-62. doi: 10.1007/BF00269667.
To establish a detailed map of the transcribed parts of the T-DNA in two octopine crown gall lines grown in suspension culture, T-DNA-derived steady-state nuclear and polysomal RNA as well as RNA synthesized in isolated nuclei purified from the crown gall tissues, was analyzed by southern blot hybridization to specific fragments of the T-region of the octopine plasmid pTi ACH5. In addition total RNA isolated from the same lines grown as callus tissue on solid agar, was analyzed for T-DNA specific transcripts. The results show that all of the T-DNA is transcribed although different segments are transcribed to significantly different extents. Roughly the same hybridization pattern was found for nuclear and polysomal poly-A+ and poly-A- RNA. The transcription pattern was found to be different for cells in the stationary phase of growth compared with actively growing cells.
为了建立悬浮培养的两种章鱼碱型冠瘿瘤系中T-DNA转录部分的详细图谱,通过与章鱼碱型质粒pTi ACH5的T区特异性片段进行Southern杂交,分析了T-DNA衍生的稳态核RNA和多聚核糖体RNA,以及从冠瘿瘤组织中纯化的分离细胞核中合成的RNA。此外,还分析了在固体琼脂上作为愈伤组织生长的相同品系中分离的总RNA的T-DNA特异性转录本。结果表明,所有T-DNA均被转录,尽管不同片段的转录程度差异显著。在核RNA和多聚核糖体的聚腺苷酸加尾(poly-A+)和非聚腺苷酸加尾(poly-A-)RNA中发现了大致相同的杂交模式。结果发现,与活跃生长的细胞相比,处于生长静止期的细胞的转录模式有所不同。