Flint S J
J Virol. 1977 Jul;23(1):44-52. doi: 10.1128/JVI.23.1.44-52.1977.
mRNA isolated from adenovirus 2-infected HeLa cells at early times during the productive cycle and from two lines of adenovirus 2-transformed rat embryo cells (F17 and T2C4) was fractionated on sucrose gradients after disaggregation. Viral mRNA species were identified by hybridization across such gradients with the separated strands of restriction endonuclease fragments of 32P-labeled DNA known to be complementary to adeovirus 2 "early" and adenovirus 2-transformed cell mRNA. mRNA transcribed from the left-hand 14% of the adenovirus 2 genome was found to comprise two species, 16 to 17S and 20 to 21S: the same sized mRNA's were present both at early times during productive infection and in the two transformed rat cell lines. Direct comparison of the sequences present in these two mRNA species by additional saturation hybridizations suggests that they are not related to one another. Three additional regions of the adenovirus 2 genome, all of which are located in the right-hand 40% of the adenovirus 2 genome, are complementary to early mRNA sequences: each of these appears to specify one major mRNA species of about 22S. Thus, five major species of adenovirus type 2 early mRNA have been identified. Two of these, copied from the left-hand 14% of the viral genome, are also present in adenovirus 2-transformed rat cells.
在生产周期早期从腺病毒2感染的HeLa细胞以及从两株腺病毒2转化的大鼠胚胎细胞系(F17和T2C4)中分离出的mRNA,在解聚后于蔗糖梯度上进行分级分离。通过与已知与腺病毒2“早期”和腺病毒2转化细胞mRNA互补的32P标记DNA的限制性内切酶片段的单链进行跨梯度杂交来鉴定病毒mRNA种类。发现从腺病毒2基因组左侧14%转录的mRNA由两种种类组成,即16至17S和20至21S:在生产性感染的早期以及在这两株转化的大鼠细胞系中都存在相同大小的mRNA。通过额外的饱和杂交对这两种mRNA种类中存在的序列进行直接比较表明它们彼此不相关。腺病毒2基因组的另外三个区域,所有这些区域都位于腺病毒2基因组右侧40%,与早期mRNA序列互补:这些区域中的每一个似乎都指定一种约22S的主要mRNA种类。因此,已鉴定出腺病毒2型早期mRNA的五种主要种类。其中两种,从病毒基因组左侧14%复制而来,也存在于腺病毒2转化的大鼠细胞中。