Sawada Y, Fujinaga K
J Virol. 1980 Dec;36(3):639-51. doi: 10.1128/JVI.36.3.639-651.1980.
Adenovirus 12 mRNA's transcribed from the transforming region were analyzed and mapped on viral DNA by the nuclease S1 gel and diazobenzyloxymethyl paper blot techniques in cells transformed of adenovirus 12 DNA and in cells early and late after lytic infection with adenovirus 12. Two initiation sites of mRNA transcription and two kinds of splicing were found in each of early regions 1A and 1B. For early region 1A mRNA's, four species were found in lytically infected cells. Three of them were commonly found in cells transformed by either HindIII-G or EcoRI-C. Cells transformed by HindIII-G contained two additional 1A transcripts, which could be the 3' portions of chimeric mRNA's of cellular-viral or viral-viral sequences. Transcription in the 1B region diverged among the above cell lines. Early after lytic infection, no appreciable amount of 1B mRNA was detected, whereas two species of mRNA's, one corresponding to protein IX mRNA and another having splicing, were found at the late stage. In cells transformed by EcoRI-C, a distinct mRNA species with splicing was observed. Cells transformed by HindIII-G contained a transcript from the leftmost part of the 1B sequence at the 5' portion of chimeric mRNA species, suggesting the presence of tandem integration of viral DNA in the cells. Other mRNA species in the 1A and 1B regions were also detected in both transformed cell lines. The results are discussed in relation to the nucleotide sequence of the HindIII-G fragment of adenovirus 12 DNA.
运用核酸酶S1凝胶和重氮苄氧基甲基纸印迹技术,对腺病毒12转化区转录的mRNA进行了分析,并将其定位在腺病毒12 DNA上,研究对象包括用腺病毒12 DNA转化的细胞以及腺病毒12裂解感染早期和晚期的细胞。在早期区域1A和1B中,均发现了mRNA转录的两个起始位点和两种剪接方式。对于早期区域1A的mRNA,在裂解感染的细胞中发现了四种类型。其中三种在被HindIII-G或EcoRI-C转化的细胞中普遍存在。被HindIII-G转化的细胞含有另外两种1A转录本,它们可能是细胞-病毒或病毒-病毒序列嵌合mRNA的3'部分。上述细胞系中1B区域的转录情况有所不同。在裂解感染早期,未检测到明显量的1B mRNA,而在晚期发现了两种mRNA,一种对应于蛋白质IX mRNA,另一种具有剪接现象。在被EcoRI-C转化的细胞中,观察到一种具有明显剪接的mRNA类型。被HindIII-G转化的细胞在嵌合mRNA类型的5'部分含有来自1B序列最左端的转录本,这表明细胞中存在病毒DNA的串联整合。在这两种转化细胞系中还检测到了1A和1B区域的其他mRNA类型。结合腺病毒12 DNA的HindIII-G片段的核苷酸序列对结果进行了讨论。