Igarashi K, Sasada R, Kurokawa T, Niiyama Y, Tsukamoto K, Sugino Y
J Virol. 1978 Oct;28(1):219-26. doi: 10.1128/JVI.28.1.219-226.1978.
By the calcium technique, intact DNA of bovine adenovirus type 3 (BAV3) was found to transform A31 cells, a clone of BALB/3T3. Transforming activity was resistant to RNase and Pronase but sensitive to DNase. The efficiency of transformation was approximately 5 to 10 foci per mug of DNA. Attempts were also made to test for transforming activity of BAV3 DNA fragments prepared with restriction endonucleases EcoRI and HindIII. The activity was found to associate exclusively with the EcoRI D fragment mapped in the region of 3.6 and 19.7 units (molecular weight, 3.9 x 10(6)). No transformation could be obtained with three HindIII fragments, J, E, and B, located at the left-hand end of the BAV3 genome. However, the enzymatic joining of J and E fragments (0 to 11.9 map units) with a ligase restored the transforming activity. These results suggest that all the genetic information of BAV3 required for transformation is located in the region between 3.6 and 11.9 units on the viral genome. Some properties of A31 cells transformed by BAV3 DNA EcoRI D fragment (TrD) and the ligated DNA of HindIII J and E fragments (TrJE), as well as those transformed by whole BAV3 DNA (Tr), were examined. As compared to untransformed A31 cells, all the transformed cell lines tested showed rapid growth, high saturation densities, and anchorage-independent growth. Moreover, they contained BAV3-specific T antigen and induced tumors in adult nude and BALB/c mice. These properties of Tr, TrD, and TrJE lines were similar to those of BAV3-transformed cells.
通过钙技术发现，牛3型腺病毒（BAV3）的完整DNA可转化BALB/3T3的一个克隆A31细胞。转化活性对核糖核酸酶和链霉蛋白酶具有抗性，但对脱氧核糖核酸酶敏感。转化效率约为每微克DNA产生5到10个集落。还尝试检测用限制性内切酶EcoRI和HindIII制备的BAV3 DNA片段的转化活性。发现该活性仅与定位在3.6和19.7单位区域（分子量为3.9×10⁶）的EcoRI D片段相关。位于BAV3基因组左端的三个HindIII片段J、E和B无法实现转化。然而，用连接酶将J和E片段（0到11.9图谱单位）进行酶促连接可恢复转化活性。这些结果表明，转化所需的BAV3所有遗传信息位于病毒基因组上3.6到11.9单位之间的区域。检测了由BAV3 DNA EcoRI D片段（TrD）和HindIII J与E片段的连接DNA（TrJE）转化的A31细胞以及由完整BAV3 DNA转化的细胞（Tr）的一些特性。与未转化的A31细胞相比，所有测试的转化细胞系均显示出生长迅速、饱和密度高以及不依赖贴壁生长。此外，它们含有BAV3特异性T抗原，并能在成年裸鼠和BALB/c小鼠中诱发肿瘤。Tr、TrD和TrJE细胞系的这些特性与BAV3转化细胞的特性相似。
