IS1550 from Mycoplasma fermentans is transposable in Escherichia coli

An insertion sequence (IS)-like element (ISMil) was previously isolated from the incognitus strain of Mycoplasma fermentans. With polymerase chain reaction primers corresponding to the left and right terminal inverted repeats of ISMil, a 1.4-kb DNA fragment was amplified from the genome of the M64 strain of M. fermentans. This DNA fragment has structural characteristics similar to those of ISMil and is designated IS1550. One copy of IS1550 encoded two considerable overlapping open reading frames (ORFs), ORF1 and ORF2. A putative translation frame-shift signal AAAAAAG (A6G) was located near the 3'-end of ORF1. This signal might cause a -1 frame-shift to form a fused product of ORF1 and ORF2 with 444 amino acids, which has a significant similarity to the putative transposase of the IS3 family. This copy of IS1550 was shown to be transposable in Escherichia coli ISM612. Its transposition caused a 1465-bp deletion immediately adjacent to the 3'-end of the element and the creation of a pair of 3-bp direct repeats flanking the element at the new insertion site. On the basis of these results, IS1550 was considered a typical transposable element.