Cloned mouse DNA fragments can replicate in a simian virus 40 T antigen-dependent system in vivo and in vitro
Cloned mouse DNA fragments can replicate in a simian virus 40 T antigen-dependent system in vivo and in vitro
复制标题
克隆的小鼠DNA片段可以在体内和体外的猿猴病毒40T抗原依赖性系统中复制
DOI：
10.1128/mcb.5.3.563-568.1985
复制
发表时间：
1985
影响因子：
5.3
通讯作者：
M. Yamada
中科院分区：
文献类型：
--
作者：
H. Ariga;Z. Tsuchihashi;M. Naruto;M. Yamada
用 BamHI 切出小鼠肝脏 DNA，克隆到 YIp5 中，其中包含 pBR322 中酿酒酵母的 URA3 基因。在分离的几个质粒中，两个质粒pMU65和pMU111可以将酿酒酵母从URA-表型转化为URA+表型，并且可以在转化体中自主复制，表明pMU65或pMU111中存在的小鼠DNA片段含有用于在酿酒酵母中复制的自主复制序列(ARS)。此外，为了确定酵母细胞中 ARS 功能与高等生物体中 ARS 功能之间的相关性，我们尝试用猿猴病毒 40 (SV40) DNA 复制系统挑战这些质粒。在所测试的两个质粒中，pMU65的EcoRI-BglII区域可以与对应于SV40 DNA起始区域的化学合成的13个核苷酸片段杂交。 pMU65（pBR322中克隆的EcoRI-BglII区域）及其亚克隆pMU65EB都可以半保守复制，在我们之前建立的体外SV40 DNA复制系统中测试这些质粒的复制活性时，DNA复制起始于EcoRI-BglII区域。此外，pMU65和pMU65EB可以在持续产生SV40 T抗原的猴Cos细胞内自主复制。这些结果表明pMU65中EcoRI-BglII区域的2.5-kilobase片段包含在SV40 DNA复制系统中复制所需的ARS。
Mouse liver DNA was cut out with BamHI and cloned into YIp5, which contained the URA3 gene of Saccharomyces cerevisiae in pBR322. Of the several plasmids isolated, two plasmids, pMU65 and pMU111, could transform S. cerevisiae from the URA- to the URA+ phenotype and could replicate autonomously within the transformant, indicating that mouse DNA fragments present in pMU65 or pMU111 contain autonomously replicating sequences (ARS) for replication in S. cerevisiae. Furthermore, to determine the correlation between ARS function in yeast cells and that in much higher organisms, we tried to challenge these plasmids with the simian virus 40 (SV40) DNA replication system. Of the two plasmids tested, the EcoRI-BglII region of pMU65 could be hybridized with a chemically synthesized 13-nucleotide fragment corresponding to the origin region of SV40 DNA. Both pMU65 (the EcoRI-BglII region cloned in pBR322) and its subclone pMU65EB could replicate semiconservatively, and initiation of DNA replication started from the EcoRI-BglII region when the replicating activity of these plasmids was tested in the in vitro SV40 DNA replication system we have established before. Furthermore, pMU65 and pMU65EB could replicate autonomously within monkey Cos cells which produce SV40 T antigen constitutively. These results show that a 2.5-kilobase fragment of the EcoRI-BglII region in pMU65 contains the ARS needed for replication in the SV40 DNA replication system.
DOI：
10.1073/pnas.80.4.1058
发表时间：
1983
影响因子：
11.1
作者：
Stohl,LL;Lambowitz,AM
通讯作者：
Lambowitz,AM
非洲爪蟾 DNA 复制起点的克隆。
DOI：
10.1073/pnas.77.9.5292
发表时间：
1980
影响因子：
11.1
作者：
Watanabe,S;Taylor,JH
通讯作者：
Taylor,JH