Conjugative transposition of Tn916: preferred targets and evidence for conjugative transfer of a single strand and for a double-stranded circular intermediate.
Transposition of conjugative transposons proceeds by excision and formation of a covalently closed circular intermediate that includes at its joint the six flanking bases from its previous host (coupling sequences). To elucidate the role of the coupling sequences in this process and to determine the sequence of targets used by Tn916, we studied its insertion into a plasmid following conjugation. The results differ from those previously observed when Tn916 was introduced by transformation. They suggest that only one specific strand of the transposon molecule is transferred during the conjugation event and that complementary strand synthesis produces a double-stranded transposon circle with no mismatches which serves as the reaction intermediate. Tn916 inserts preferentially at specific sites and the same targets are used when Tn916 comes from donors with different coupling sequences. An analysis of the sequences of preferred targets is presented.
Department of Microbiology and Immunology, Emory University Health Sciences Center, Atlanta, Georgia 30322.
J R
JR
Scott
F
F
Bringel
D
D
Marra
G
G
Van Alstine
C K
CK
Rudy
D001412:Bacillus subtilis / Q000235:genetics; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002702:Chloramphenicol Resistance / Q000235:genetics; D003227:Conjugation, Genetic; D003433:Crosses, Genetic; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004270:DNA, Circular / Q000235:genetics; D004277:DNA, Single-Stranded / Q000235:genetics; D013293:Enterococcus faecalis / Q000235:genetics; D018014:Gene Transfer Techniques; D006094:Gram-Positive Bacteria / Q000235:genetics*; D013294:Lactococcus lactis / Q000235:genetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004270:DNA, Circular; D004277:DNA, Single-Stranded; D015500:Chloramphenicol O-Acetyltransferase