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The integrative conjugative element (ICE) Tn 5253  of  Streptococcus pneumoniae , conferring resistance to tetracycline and chloramphenicol, was found integrated at a 83-bp specific target site ( att B) located in the  rbgA  gene of the pneumococcal chromosome. PCR analysis of Tn 5253 -carrying strains showed evidence of precise excision of Tn 5253  from the pneumococcal chromosome with production of (i) circular forms of the ICE in which the ends were joined by a 84-bp sequence ( att Tn), and (ii) reconstituted chromosomal  att B. When integrated into the chromosome, Tn 5253  was flanked by  att L, identical to  att B, and  att R, identical to  att Tn. Circular forms of Tn 5253  were present at a concentration of 3.8 × 10 -4  copies per chromosome, whereas reconstituted  att B sites were at 3.0 × 10 -4  copies per chromosome. Deletion of  int-xis  of Tn 5253  abolished production of circular forms (<7.1 × 10 -6  copies per chromosome) and was associated to the lack of Tn 5253  conjugal transfer suggesting, as expected, that Tn 5253  circular form acts as a conjugation intermediate.

Excision and Circularization of Integrative Conjugative Element Tn5253 of Streptococcus pneumoniae