Hecht D W, Malamy M H
Department of Molecular Biology and Microbiology, Tufts University Health Sciences Campus, Boston, Massachusetts 02111.
J Bacteriol. 1989 Jul;171(7):3603-8. doi: 10.1128/jb.171.7.3603-3608.1989.
Conjugal transposons play an important role in the dissemination of antibiotic resistance determinants in the streptococci and have been postulated to exist in Bacteroides fragilis. To investigate the presence of conjugal transposons in B. fragilis, we employed a Tra- derivative of the transfer factor pBFTM10 contained in the chimeric plasmid pGAT400 delta BglII. We attempted to restore transferability to this plasmid from a series of transconjugants generated by crossing B. fragilis TMP230 containing the TET transfer factor with B. fragilis TM4000, a standard recipient. Transconjugant TM4.2321 transferred pGAT400 delta BglII to Escherichia coli HB101 at almost the same frequency as did the Tra+ parental plasmid, pGAT400. Analysis of the transferred plasmids revealed the presence of 9.6 kilobases of additional DNA in every case but at different positions in independent isolates. The presence of this DNA, designated Tn4399, allowed the pGAT400 delta BglII derivatives to retransfer from the TM4000 background to B. fragilis or E. coli recipients. DNA hybridization studies demonstrated the presence of one copy of Tn4399 in TMP230 and three copies at new sites in TM4.2321. Tn4399 is a new B. fragilis transposon with unique transfer properties that may play a role in the dissemination of drug resistance genes. It differs from previously described conjugal transposons by its ability to mobilize nonconjugal plasmids in cis.
接合转座子在链球菌中抗生素抗性决定因素的传播中起重要作用,并且据推测存在于脆弱拟杆菌中。为了研究脆弱拟杆菌中接合转座子的存在情况,我们使用了嵌合质粒pGAT400 delta BglII中所含转移因子pBFTM10的一个无Tra基因的衍生物。我们试图从一系列由含有TET转移因子的脆弱拟杆菌TMP230与标准受体脆弱拟杆菌TM4000杂交产生的转接合子中恢复该质粒的转移性。转接合子TM4.2321将pGAT400 delta BglII转移至大肠杆菌HB101的频率几乎与Tra+亲本质粒pGAT400相同。对转移质粒的分析显示,在每种情况下均存在9.6千碱基的额外DNA,但在独立分离株中的位置不同。这种被命名为Tn4399的DNA的存在使pGAT400 delta BglII衍生物能够从TM4000背景重新转移至脆弱拟杆菌或大肠杆菌受体。DNA杂交研究表明,Tn4399在TMP230中有一个拷贝,在TM4.2321的新位点有三个拷贝。Tn4399是一种新的脆弱拟杆菌转座子,具有独特的转移特性,可能在耐药基因的传播中起作用。它与先前描述的接合转座子的不同之处在于其能够顺式动员非接合性质粒。