Francia M V, García Lobo J M
Departamento de Biologia Molecular, Facultad de Medicina, Universidad de Cantabria, Santander, Spain.
J Bacteriol. 1996 Feb;178(3):894-8. doi: 10.1128/jb.178.3.894-898.1996.
A replication-thermosensitive, pSC101-derived plasmid containing the int gene and RHS-2 from the integron in Tn21 and a kanamycin resistance marker has been constructed and used to obtain Tn21 integrase (Int21)-mediated plasmid integration in the Escherichia coli chromosome. Colonies carrying an integrated plasmid were obtained after growth at 42 degrees C. Southern hybridization and PCR experiments indicated that they contained the plasmid specifically integrated through the RHS into different positions in the E. coli chromosome. Nucleotide sequence determination of the plasmid-chromosome junctions showed that integration sites in the chromosome were pentanucleotides with the sequence described for Int21 secondary sites.
构建了一种源自pSC101的复制温度敏感型质粒，其含有来自Tn21中整合子的int基因和RHS-2以及卡那霉素抗性标记，并用于在大肠杆菌染色体中获得Tn21整合酶（Int21）介导的质粒整合。在42℃生长后获得了携带整合质粒的菌落。Southern杂交和PCR实验表明，它们含有通过RHS特异性整合到大肠杆菌染色体不同位置的质粒。质粒-染色体连接点的核苷酸序列测定表明，染色体中的整合位点是具有Int21二级位点所述序列的五核苷酸。
