Universität Bielefeld, Fakultät für Biologie (Genetik), D-4800 Bielefeld 1, FRG.
EMBO J. 1985 May;4(5):1119-24. doi: 10.1002/j.1460-2075.1985.tb03748.x.
A genomic DNA fragment containing a soybean heat shock gene (hs6871) was inserted into the T-DNA region of the Agrobacterium tumefaciens pTiC58 plasmid. A strain carrying the modified Ti plasmid was used to incite tumors in sunflower hypocotyls. The expression of the heat shock gene was investigated by Northern blot analysis of RNA and S1 nuclease mapping of the transcriptional start site. Heat shock-induced poly(A) mRNA was detected in tumor tissue only after incubation at 40 degrees C (heat shock) not at the normal growth temperature (28 degrees C). Transcripts from hs6871 are faithfully initiated in sunflower, starting at the same site on the DNA as in soybean. The low level of transcripts initiating correctly on hs6871 in sunflower is consistent with a general tissue-specific reduction in the expression of partially homologous native heat shock genes in sunflower tumors.
一个包含大豆热激基因(hs6871)的基因组 DNA 片段被插入到农杆菌 Ti 质粒的 T-DNA 区。携带修饰过的 Ti 质粒的菌株被用于诱导向日葵下胚轴的肿瘤。通过 Northern 印迹分析 RNA 和 S1 核酸酶转录起始位点作图来研究热激基因的表达。只有在 40 度(热激)而不是在正常生长温度(28 度)下孵育后,才能在肿瘤组织中检测到热激诱导的多(A)mRNA。hs6871 的转录物在向日葵中被忠实地起始,从 DNA 上与大豆中相同的位点开始。在向日葵中正确起始 hs6871 的转录物水平较低,这与在向日葵肿瘤中部分同源的天然热激基因的表达在组织上普遍降低相一致。