Rajagopalan S, Chinnadurai G
Nucleic Acids Res. 1979 Nov 10;7(5):1163-74. doi: 10.1093/nar/7.5.1163.
We have identified a novel subgenomic viral DNA in KB cells infected with adenovirus 2 (Ad2) under high multiplicities of infection. KB cells were infected with Ad2 at multiplicities of infection greater than 100 PFU/cell. 32P-labeled viral DNA was selectively extracted by a modification of the method of Hirt (8) from the infected cells and analyzed by electrophoresis on agarose gels. In addition to full-length DNA (33 to 23 x 10(6) daltons), a unique subgenomic DNA species of about 12 to 13% (2.6 x 10(6) daltons) of full-length DNA in size was found in the infected cells. This subgenomic DNA was found to be double stranded and was not packaged inside the virus particles. This DNA could be isolated in large amounts (30 to 50% of total viral DNA) from infected cells. When cleaved with restriction endonuclease KpnI, the subgenomic DNA yielded two fragments, each corresponding to about 6% and 7% of the full-length genome in size.
我们在高感染复数下感染腺病毒2（Ad2）的KB细胞中鉴定出一种新型亚基因组病毒DNA。KB细胞以大于100 PFU/细胞的感染复数感染Ad2。通过对Hirt（8）方法的改进，从感染细胞中选择性提取32P标记的病毒DNA，并在琼脂糖凝胶上进行电泳分析。除了全长DNA（33至23×10⁶道尔顿）外，在感染细胞中还发现了一种独特的亚基因组DNA，其大小约为全长DNA的12%至13%（2.6×10⁶道尔顿）。这种亚基因组DNA被发现是双链的，且未包装在病毒颗粒内。这种DNA可以从感染细胞中大量分离（占总病毒DNA的30%至50%）。用限制性内切酶KpnI切割时，亚基因组DNA产生两个片段，每个片段的大小约对应全长基因组的6%和7%。
