Wróbel B, Wegrzyn G
Department of Molecular Biology, University of Gdańsk, Poland.
J Biotechnol. 1997 Dec 3;58(3):205-8. doi: 10.1016/s0168-1656(97)00148-x.
Amino acid starvation of Escherichia coli relA mutants was previously proposed as a method for efficient amplification of plasmids bearing origin of replication derived from ColE1-type plasmids and bacteriophage lambda. Here we demonstrate that plasmids derived from pSC101 replicon can be amplified in E. col relA+ and relA- strains during amino acid limitation but not during amino acid starvation. The amplification efficiency is dependent on temperature (37 degrees C was found to be the optimal temperature). Under optimal conditions, up to 13-fold amplification of a pSC101-derived plasmid may be achieved.
以前有人提出,使大肠杆菌relA突变体处于氨基酸饥饿状态是一种有效扩增携带源自ColE1型质粒和噬菌体λ复制起点的质粒的方法。在此我们证明,源自pSC101复制子的质粒在氨基酸限制期间可在大肠杆菌relA+和relA-菌株中扩增,但在氨基酸饥饿期间则不能。扩增效率取决于温度(发现37摄氏度是最佳温度)。在最佳条件下,源自pSC101的质粒可实现高达13倍的扩增。