NEW COSMID VECTORS DEVELOPED FOR EUKARYOTIC DNA CLONING
NEW COSMID VECTORS DEVELOPED FOR EUKARYOTIC DNA CLONING
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DOI：
10.1016/0378-1119(84)90143-4
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发表时间：
1984-01-01
期刊：
影响因子：
3.5
通讯作者：
HASHIMOTOGOTOH, T
中科院分区：
文献类型：
--
作者：
BRADY, G;JANTZEN, HM;HASHIMOTOGOTOH, T
构建了一系列适合克隆大片段DNA的ColE 1和pSC 101粘粒载体。它们都含有一个BamHI位点，允许克隆Sau 3A、MboI、BglII、BclI和BamHI产生的片段。这些载体具有以下特点：（1.7-3.4 kb [内切酶]）; BamHI克隆位点的侧翼是限制酶位点，能够直接克隆未分级的插入DNA而不产生多个插入或载体连接产物; 2载体（pHSG 272和pHSG 274）含有在原核和真核细胞中均可选择的杂合Tn 5 KmR/G418 R基因，使它们适于将DNA转移到真核细胞中，以及在所述的其它载体中可获得的不同原核选择标记促进了从真核细胞转移的DNA序列的粘粒拯救：CmR、ApR、KmR（pHSG 429）、CmR（pHSG 439）、大肠杆菌素E1免疫（pHSG 250），粘粒pHSG 272被成功地用于构建基于BPVI [牛乳头瘤病毒]复制子的穿梭载体。
A series of ColE1 and pSC101 cosmid vectors were constructed suitable for cloning large stretches of DNA. All contain a single BamHI site allowing cloning of Sau3A, MboI, BglII, BclI and BamHI-generated fragments. These vectors have the following characteristics: they are relatively small (1.7-3.4 kb [kilobase]); the BamHI cloning site is flanked by restriction enzyme sites enabling direct cloning of unfractionated insert DNA without generating multiple insert or vector ligation products; 2 vectors (pHSG272 and pHSG274) contain a hybrid Tn5 KmR/G418R gene which is selectable in both prokaryotic and eukaryotic cells, making them suitable for transferring DNA into eukaryotic cells, and the different prokaryotic selectable markers available in the other vectors described facilitate cosmid rescue of the transferred DNA sequences from the eukaryotic cell: CmR, ApR, KmR, (pHSG429), CmR, (pHSG439), colicin E1 immunity (pHSG250), the cosmid pHSG272 was used successfully to construct a shuttle vector based on the BPVI [bovine papilloma virus] replicon.