Brady G, Jantzen H M, Bernard H U, Brown R, Schütz G, Hashimoto-Gotoh T
Gene. 1984 Feb;27(2):223-32. doi: 10.1016/0378-1119(84)90143-4.
A series of ColE1 and pSC101 cosmid vectors have been 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: (i) they are relatively small (1.7-3.4 kb); (ii) 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 [ Ish - Horowitz and Burke, Nucl . Acids Res. 9 (1981) 2989-2998]; (iii) two 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 (iv) 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 ), (v) the cosmid pHSG272 was used successfully to construct a shuttle vector based on the BPVI replicon [ Matthias et al., EMBO J. 2 (1983) 1487-1492].
已构建了一系列适合克隆大片段DNA的ColE1和pSC101黏粒载体。所有载体都含有一个单一的BamHI位点，可用于克隆Sau3A、MboI、BglII、BclI和BamHI产生的片段。这些载体具有以下特点：（i）它们相对较小（1.7 - 3.4 kb）；（ii）BamHI克隆位点两侧是限制酶位点，可直接克隆未分级的插入DNA，而不会产生多个插入或载体连接产物[Ish - Horowitz和Burke，《核酸研究》9（1981）2989 - 2998]；（iii）两个载体（pHSG272和pHSG274）含有一个杂交的Tn5 KmR/G418R基因，该基因在原核细胞和真核细胞中均可选择，使其适合将DNA转移到真核细胞中，并且（iv）其他所述载体中可用的不同原核选择标记有助于从真核细胞中黏粒拯救转移的DNA序列：CmR、ApR、KmR（pHSG429）、CmR（pHSG439）、大肠杆菌素E1免疫（pHSG250），（v）黏粒pHSG272已成功用于构建基于BPVI复制子的穿梭载体[Matthias等人，《欧洲分子生物学组织杂志》2（1983）1487 - 1492]。
