一种用于快速鉴定大肠杆菌HB101细胞中重组pAT153质粒的简化方法。

A simplified procedure for the rapid identification of recombinant pAT153 plasmids in Escherichia coli HB101 cells.

作者信息

Dion M, Yelle J, Hamelin C

出版信息

Anal Biochem. 1986 Apr;154(1):209-12. doi: 10.1016/0003-2697(86)90517-8.

Abstract

Insertion of foreign DNA into the unique HindIII site of the high copy number plasmid pAT153 reduces but does not completely abolish the resistance of Escherichia coli HB101 cells to tetracycline. Recombinant DNA-containing colonies could then be phenotypically differentiated from non-recombinant ones by their smaller size on nutrient agar plates with ampicillin and tetracycline at a final concentration of 50 and 4 micrograms/ml, respectively. A wide variety of human cytomegalovirus DNA fragments have been found in pAT153 molecules propagated by the ampicillin-resistant tetracycline-sensitive bacteria selected.

摘要

将外源DNA插入高拷贝数质粒pAT153的唯一HindIII位点会降低但不会完全消除大肠杆菌HB101细胞对四环素的抗性。然后，通过在含有终浓度分别为50微克/毫升和4微克/毫升氨苄青霉素和四环素的营养琼脂平板上，含重组DNA的菌落尺寸较小，可从非重组菌落中进行表型区分。在通过选择的对氨苄青霉素抗性、对四环素敏感的细菌繁殖的pAT153分子中发现了多种人巨细胞病毒DNA片段。