COLE1-TYPE VECTORS WITH FULLY REPRESSIBLE REPLICATION
COLE1-TYPE VECTORS WITH FULLY REPRESSIBLE REPLICATION
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DOI：
10.1016/0378-1119(91)90508-9
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发表时间：
1991-08-30
期刊：
影响因子：
3.5
通讯作者：
BOUCHE, JP
中科院分区：
文献类型：
--
作者：
GIL, D;BOUCHE, JP
我们从pBR322中构建了两个克隆载体pAM34和pAM35，其中复制引物RNA的转录受lacZpo启动子/操纵子的控制。这些载体包含一个克隆盒，两侧有强大的转录终止子。它们的不同之处在于LacI(Q)基因的存在(PAM34)或缺失(PAM35)。在抑制子LaCI(Q)的存在下，复制完全依赖于诱导剂的添加。这一特征允许暂时维持这些质粒，构建其中载体衍生物稳定整合到染色体中的菌株，以及恢复克隆片段附近的核苷酸序列。整合的质粒的复制可以被调节，以匹配在没有功能起始物的情况下细胞生长所需的染色体复制起始率。
We have constructed two cloning vectors, pAM34 and pAM35, derived from pBR322, in which transcription of the replication primer RNA is under control of the lacZpo promoter/operator. These vectors contain a cloning cassette flanked by strong transcriptional terminators. They differ from each other by the presence (pAM34) or absence (pAM35) of gene lacI(q). In the presence of repressor LacI(q), replication is entirely dependent upon the addition of inducer. This feature allows the temporary maintenance of these plasmids, the construction of strains in which vector derivatives are stably integrated into the chromosome, and the recovery of nucleotide sequences adjacent to cloned fragments. Replication from the integrated plasmid can be adjusted to match the chromosome replication initiation rate required for cell growth in the absence of a functional origin, oriC.