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Formation of an inverted repeat junction in the transposition of insertion sequence ISLC3 isolated from Lactobacillus casei

Institute of Molecular Medicine and Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan, ROC

Correspondence
Thy-Hou Lin
thlin{at}life.nthu.edu.tw

An insertion sequence, ISLC3, of 1351 bp has been isolated from Lactobacillus casei. Formation of IS circles containing a 3 bp spacer (complete junction) or deletion of 25 bp at the left inverted repeat (IRL) between the abutted IS ends of the ISLC3 junction region (deleted junction) was also discovered in the lactobacilli and Escherichia coli system studied. We found that the promoter formed by the complete junction P_jun was more active than that formed by the 25 bp deleted junction P_djun or the indigenous promoter P_IRL. The corresponding transcription start sites for both promoter P_jun and P_IRL as well as P_djun were subsequently determined using a primer extension assay. The activity of transposase OrfAB of ISLC3 was also assayed using an in vitro system. It was found that this transposase preferred to cleave a single DNA strand at the IRR over the IRL end in the transposition process, suggesting that attack of one end by the other was oriented from IRR to IRL.