Construction and Use of Derivatives of Transposon Tn 4001 That Function in Mycoplasma pulmonis and Mycoplasma arthritidis

Abstract

ABSTRACT Previous attempts to introduce transposon Tn 4001 into Mycoplasma pulmonis and Mycoplasma arthritidis have not been successful, possibly due to functional failure of the transposon's gentamicin resistance determinant. Tn 4001C and Tn 4001T were constructed, respectively, by insertion of a chloramphenicol acetyltransferase gene and the tetM tetracycline resistance determinant into Tn 4001 . Both Tn 4001C and Tn 4001T transposed in M. pulmonis , and Tn 4001T transposed in M. arthritidis . The incorporation of a Tn 4001T derivative that contained lacZ into either Mycoplasma species resulted in transformants with readily detectable LacZ activity. Tn 4001T may be of general utility for use as a mycoplasma cloning vehicle because tetM functions in all species of Mycoplasma examined thus far.