Naglich J G, Andrews R E
Department of Microbiology, Iowa State University, Ames 50011.
Plasmid. 1988 Mar;19(2):84-93. doi: 10.1016/0147-619x(88)90047-9.
The conjugative Streptococcus faecalis transposon Tn916 was introduced into Bacillus thuringiensis subsp. israelensis by filter matings with S. faecalis. B. thuringiensis transconjugants resistant to tetracycline (Tetr) were detected at a frequency of approximately 7.0 X 10(-7) per recipient cell during filter matings, whereas transfer of Tn916 was not observed in broth matings. The Tetr phenotype in subsp. israelensis was stable in the absence of antibiotic selection. Southern hybridization analysis revealed that Tn916 had inserted into several different sites on the B. thuringiensis subsp. israelensis chromosome but insertion into plasmid DNA was not observed. Movement of Tn916 was demonstrated when Tetr B. thuringiensis transconjugants were mated with isogenic recipients. Southern hybridizations, however, showed that the resulting Tetr isolates contained Tn916 junction fragments that were nearly identical to the donor, suggesting that this movement resulted from transfer of chromosomal DNA from donor to recipient or from a fusion of mating cells, rather than conjugative transposition of the Tn element.
通过与粪肠球菌进行滤膜杂交,将接合型粪肠球菌转座子Tn916导入苏云金芽孢杆菌以色列亚种。在滤膜杂交过程中,每受体细胞中检测到对四环素(Tetr)具有抗性的苏云金芽孢杆菌转接合子的频率约为7.0×10(-7),而在肉汤杂交中未观察到Tn916的转移。在没有抗生素选择的情况下,以色列亚种中的Tetr表型是稳定的。Southern杂交分析表明,Tn916已插入苏云金芽孢杆菌以色列亚种染色体上的几个不同位点,但未观察到插入质粒DNA。当Tetr苏云金芽孢杆菌转接合子与同基因受体杂交时,证明了Tn916的移动。然而,Southern杂交表明,产生的Tetr分离株含有与供体几乎相同的Tn916连接片段,这表明这种移动是由于染色体DNA从供体转移到受体或由于杂交细胞的融合,而不是Tn元件的接合转座。