Gene integration in the Escherichia coli chromosome mediated by Tn21 integrase (Int21)

被引:14
作者
Francia, MV [1 ]
Lobo, JMG [1 ]
机构
[1] UNIV CANTABRIA,FAC MED,DEPT BIOL MOLEC,E-39005 SANTANDER,SPAIN
关键词
D O I
10.1128/jb.178.3.894-898.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A replication-thermosensitive, pSC101-derived plasmid containing the inf 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.
引用
收藏
页码:894 / 898
页数:5
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