Infection of eucaryotic cells by helper-independent recombinant adenoviruses: early region 1 is not obligatory for integration of viral DNA
Infection of eucaryotic cells by helper-independent recombinant adenoviruses: early region 1 is not obligatory for integration of viral DNA
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不依赖辅助细胞的重组腺病毒感染真核细胞：早期区域 1 对于病毒 DNA 的整合不是必需的
DOI：
10.1128/jvi.50.2.606-614.1984
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发表时间：
1984
影响因子：
5.4
通讯作者：
Cold Spring Hlrhbor
中科院分区：
文献类型：
--
作者：
Kevin Van Doren;Douglas Hanahan;Y. Gluzman;Cold Spring Harbor Laborator;Cold Spring Hlrhbor
通过将新霉素抗性的杂交基因插入到辅助子非依赖型腺病毒载体Delta E1/X中，构建了携带可选择耐药标记的重组病毒基因组。该杂交基因由编码来自TN5的氨基糖苷3‘-磷酸转移酶II的序列组成，受猴病毒40早期启动子的控制，并使哺乳动物细胞对新霉素类似物G-418产生抗性。腺病毒早期区1的大部分缺失于增量E_1/X(核苷酸455~3330)，重组病毒基因组在其位置携带杂交基因。将Delta E1/X的大、小XbaI片段分别连接到杂合基因上，并将其导入293细胞。分离单个斑块，然后在293细胞中传代，以产生病毒库。重组病毒有效地使培养的大鼠(Rat2)和猿猴(CV1)细胞对G-418产生抗性。对G-418具有抗性的克隆细胞系含有整合到宿主细胞基因组中的病毒DNA，表明早期区域1对病毒基因组的整合不是必不可少的。Southern转移实验表明：(I)整合位点在宿主基因组中并不是唯一的；(Ii)转化的CV1细胞通常含有单拷贝的全长病毒基因组，与侵染病毒共线；(Iii)转化的Rat2细胞株一般包含一到多个拷贝的全长病毒基因组与侵染的病毒DNA共生；以及(Iv)这五个转化的Rat2细胞系中的三个包含重复的病毒DNA序列，其中病毒基因组的左右两端相互连接。
Recombinant viral genomes carrying a selectable drug resistance marker have been constructed by insertion of a hybrid gene for neomycin resistance into the helper-independent adenovirus vector, delta E1/X. The hybrid gene consists of sequences coding for the aminoglycoside 3'-phosphotransferase II from Tn5, under the control of the simian virus 40 early promoter, and renders mammalian cells resistant to the neomycin analog, G-418. Most of adenovirus early region 1 is deleted from delta E1/X (nucleotides 455 to 3330), and recombinant viral genomes carry the hybrid gene in its place. The large and small XbaI fragments of delta E1/X were ligated to the hybrid gene, and the mixture was transfected into 293 cells. Single plaques were isolated and subsequently passaged in 293 cells to produce virus stocks. The recombinant viruses efficiently rendered cultured rat (Rat2) and simian (CV1) cells resistant to G-418. Cloned cell lines selected for resistance to G-418 contained viral DNA integrated into the host cell genome, demonstrating that early region 1 is not essential for integration of the viral genome. Southern transfer experiments revealed that (i) the sites of integration in the host genome were not unique; (ii) in general, transformed CV1 cell lines contained single-copy, full-length viral genomes, colinear with the infecting virus; (iii) transformed Rat2 cell lines generally contained one to several copies of full-length viral genomes integrated colinearly with the infecting viral DNA; and (iv) three of these five lines of transformed Rat2 cell lines contained tandemly repeated viral DNA sequences in which the right and left ends of the viral genome were joined to each other.
具有冷敏感宿主范围表型的 5 型腺病毒突变体的分离：腺病毒转化维持功能的遗传证据。
影响因子：
3.7
作者：
Ho,YS;Galos,R;Williams,J
通讯作者：
Williams,J
影响因子：
56.9
作者：
C. Miller;H. Temin
通讯作者：
C. Miller;H. Temin
5 型腺病毒宿主范围突变体的冷敏感转化表达。
DOI：
10.1073/pnas.80.5.1352
发表时间：
1983
影响因子：
11.1
作者：
Babiss,LE;Ginsberg,HS;Fisher,PB
通讯作者：
Fisher,PB