Transcription map for adenovirus type 12 DNA
Transcription map for adenovirus type 12 DNA
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12 型腺病毒 DNA 转录图谱
DOI：
10.1128/jvi.28.1.227-239.1978
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发表时间：
1978
影响因子：
5.4
通讯作者：
S. Mak
中科院分区：
文献类型：
--
作者：
J. Smiley;S. Mak
12型腺病毒基因组的编码l-和r-链特异性胞质RNA的区域通过以下程序作图。将病毒基因组的放射性、完整、分离的互补链与饱和量的未标记晚期细胞质RNA杂交。然后通过杂交体的S1核酸酶消化纯化与RNA互补的每条DNA链的片段。每条链的编码区的排列由这些探针与用EcoRI、BamHI和HindIII消化产生的未标记病毒DNA片段的杂交模式推断。所得到的图谱与2型腺病毒的图谱相似，如果不相同的话。通过将标记的早期细胞质RNA与未标记的DNA片段和未标记的特异性片段的互补链杂交，在图上定位在早期表达的晚期细胞质RNA序列的子集。早期细胞质RNA与EcoRI-C和BamHI-B的r-链和BamHI-E的l-链杂交。还观察到与BamHI-C的杂交。在早期和晚期测量与单个限制性片段互补的细胞质RNA积累的相对速率。在感染早期，出现在细胞质中的大部分病毒RNA来自基因组的分子末端。后来（感染后24至26小时），大多数新标记的细胞质RNA转录自基因组上25至60个图单位之间的DNA序列映射。
The regions of the adenovirus type 12 genome which encode l- and r-strand-specific cytoplasmic RNA were mapped by the following procedure. Radioactive, intact, separated complementary strands of the viral genome were hybridized to saturating amounts of unlabeled late cytoplasmic RNA. The segments of each DNA strand complementary to the RNA were then purified by S1 nuclease digestion of the hybrids. The arrangement of the coding regions of each strand was deduced from the pattern of hybridization of these probes to unlabeled viral DNA fragments produced by digestion with EcoRI, BamHI, and HindIII.. The resulting map is similar, if not identical, to that of adenovirus type 2. The subset of the late cytoplasmic RNA sequences which are expressed at early times were located on the map by hybridizing labeled, early cytoplasmic RNA to both unlabeled DNA fragments and unlabeled complementary strands of specific fragments. Early cytoplasmic RNA hybridized to the r-strand to EcoRI-C and BamHI-B and to the l-strand of BamHI-E. Hybridization to BamHI-C was also observed. The relative rates of accumulation of cytoplasmic RNA complementary to individual restriction fragments was measured at both early and late times. Early during infection, most of the viral RNA appearing in the cytoplasm was derived from the molecular ends of the genome. Later (24 to 26 h postinfection) the majority of the newly labeled cytoplasmic RNA was transcribed from DNA sequences mapping between 25 and 60 map units on the genome.