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氯霉素乙酰转移酶基因在早期腺病毒C亚组启动子控制下在人细胞中的表达:来自其他亚组的E1A基因产物对基因表达的影响。

Expression of the chloramphenicol acetyl transferase gene in human cells under the control of early adenovirus subgroup C promoters: effect of E1A gene products from other subgroups on gene expression.

作者信息

Leite J P, Niel C, D'Halluin J C

出版信息

Gene. 1986;41(2-3):207-15. doi: 10.1016/0378-1119(86)90100-9.

DOI:10.1016/0378-1119(86)90100-9
PMID:3458651
Abstract

A hierarchy of dominance has been observed in HeLa cells co-infected with two serotypes of adenovirus belonging to different subgroups. DNA replication and late protein synthesis of one serotype are inhibited by those of the other. The degree of inhibitory effect has the following decreasing order: adenovirus type 3 (Ad3) and Ad7 (subgroup B), Ad9 (D), Ad4 (E), Ad12 (A), Ad2 and Ad5 (C) [Delsert and D'Halluin, Virus Res. 1 (1984) 365-380]. HeLa cells were first transfected with recombinant plasmids carrying Ad5 E2A or E3 promoters fused to the chloramphenicol acetyl transferase gene (cat), and then infected with human Ad belonging to different subgroups. All the serotypes tested were found to be able to stimulate both E2A and E3 promoters. When HeLa cells were co-transfected with either of the previous plasmids, plus a second plasmid carrying the Ad3 E1A region, the same stimulatory effect was observed. However, an inhibitory effect on Ad5 E2A and E3 promoters seemed to occur when both Ad2 E1A (subgroup C) and Ad3 E1A (subgroup B) genes were present together. To determine which one of the early products was responsible for the observed repression effect, and to assign the target on the genome of subgroup C Ad, a plasmid was constructed in which the sequences at the 5' end of the Ad2 E1A region were fused to the structural sequences of the cat gene. In HeLa cells transfected with this plasmid, CAT activity was significantly increased after co-transfection with a plasmid carrying the Ad2 E1A region, but decreased with a plasmid carrying the Ad3 E1A region.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在感染了属于不同亚组的两种血清型腺病毒的HeLa细胞中观察到了一种优势等级。一种血清型的DNA复制和晚期蛋白质合成受到另一种血清型的抑制。抑制作用的程度按以下递减顺序排列:3型腺病毒(Ad3)和7型腺病毒(Ad7,B亚组)、9型腺病毒(Ad9,D亚组)、4型腺病毒(Ad4,E亚组)、12型腺病毒(Ad12,A亚组)、2型腺病毒和5型腺病毒(Ad2和Ad5,C亚组)[德尔塞特和德哈兰,《病毒研究》1(1984年)365 - 380页]。HeLa细胞首先用携带与氯霉素乙酰转移酶基因(cat)融合的Ad5 E2A或E3启动子的重组质粒进行转染,然后用属于不同亚组的人腺病毒进行感染。发现所有测试的血清型都能够刺激E2A和E3启动子。当HeLa细胞用上述任何一种质粒加上携带Ad3 E1A区域的第二种质粒进行共转染时,观察到了相同的刺激作用。然而,当Ad2 E1A(C亚组)和Ad3 E1A(B亚组)基因同时存在时,似乎对Ad5 E2A和E3启动子产生了抑制作用。为了确定哪种早期产物负责观察到的抑制作用,并确定C亚组腺病毒基因组上的靶点,构建了一种质粒,其中Ad2 E1A区域5'端的序列与cat基因的结构序列融合。在用该质粒转染的HeLa细胞中,与携带Ad2 E1A区域的质粒共转染后,CAT活性显著增加,但与携带Ad3 E1A区域的质粒共转染后则降低。(摘要截短于250字)

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Proc Natl Acad Sci U S A. 1986 Mar;83(6):1598-1602. doi: 10.1073/pnas.83.6.1598.

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