Cowie A, de Villiers J, Kamen R
Mol Cell Biol. 1986 Dec;6(12):4344-52. doi: 10.1128/mcb.6.12.4344-4352.1986.
We have identified a putative DNA-binding domain in polyomavirus large T antigen. Mutations introduced into the gene between amino acids 290 and 310 resulted in proteins that no longer bound to the high-affinity binding sites on the polyomavirus genome, showed no detectable nonspecific DNA binding, and were not able to initiate DNA replication from the viral origin. These mutant T antigen genes were introduced into rat embryo fibroblasts together with the neomycin resistance gene to allow selection for growth in the presence of G418. All the mutations tested facilitated the establishment of these cells in long-term culture at an efficiency indistinguishable from that of the wild-type protein.
我们已经在多瘤病毒大T抗原中鉴定出一个假定的DNA结合结构域。在氨基酸290和310之间的基因中引入的突变导致蛋白质不再与多瘤病毒基因组上的高亲和力结合位点结合，未显示可检测到的非特异性DNA结合，并且无法从病毒起源处启动DNA复制。这些突变的T抗原基因与新霉素抗性基因一起被导入大鼠胚胎成纤维细胞，以便在存在G418的情况下进行生长选择。所有测试的突变都促进了这些细胞在长期培养中的建立，其效率与野生型蛋白质的效率没有区别。
