Department of Biochemistry, University of Washington, Seattle, Washington 98195.
Proc Natl Acad Sci U S A. 1980 May;77(5):2666-70. doi: 10.1073/pnas.77.5.2666.
In order to detect proteins that may be produced in crown gall tumors as a result of expression of incorporated Agrobacterium tumefaciens Ti plasmid DNA (T-DNA), we have isolated mRNA complementary to T-DNA and translated this in a protein-synthesizing system derived from wheat germ. mRNA prepared from cultured E1 tumor from Nicotiana tabacum hybridized with HindIII fragment 1 sequences of T-DNA immobilized on cellulose nitrate filters. Two proteins of 30,000 and 16,500 M(r) were produced when this selected RNA was released and translated. Other tumor lines from N. tabacum were investigated, and a protein of slightly less than 30,000 M(r) was encoded by HindIII fragment 1 sequences of 15955/01 tumor. No products were observed for 15955/1 tumor line, which differs from E1/B6-806 and 15955/01 in that it does not produce octopine. mRNA species of each of the tumor lines hybridized to Bst I fragment 8 sequences of T-DNA and produced a common protein of 15,000 M(r). Because this protein is derived from the region of the T-DNA that is conserved in octopine- and nopaline-type crown gall tumors, it may play a role in oncogenicity.
为了检测由于整合的根癌农杆菌 Ti 质粒 DNA(T-DNA)表达而可能在冠瘿瘤中产生的蛋白质,我们已经分离了与 T-DNA 互补的 mRNA,并在源自小麦胚芽的蛋白质合成系统中翻译了它。从培养的烟草杂种 E1 肿瘤中制备的 mRNA 与固定在硝酸纤维素滤器上的 T-DNA HindIII 片段 1 序列杂交。当释放并翻译此选择的 RNA 时,产生了 30,000 和 16,500 Mr 的两种蛋白质。还研究了来自烟草的其他肿瘤系,并且由 15955/01 肿瘤的 HindIII 片段 1 序列编码的略微小于 30,000 Mr 的蛋白质。对于 15955/1 肿瘤系未观察到产物,该肿瘤系与 E1/B6-806 和 15955/01 不同,因为它不产生 octopine。每个肿瘤系的 mRNA 与 T-DNA 的 Bst I 片段 8 序列杂交,并产生了 15,000 Mr 的共同蛋白质。由于该蛋白质来源于 octopine 和 nopaline 型冠瘿瘤中保守的 T-DNA 区域,因此它可能在致癌性中起作用。