TY - JOUR
T1 - Orientation and sequence analysis of right ends and target sites of bacteriophage Mu and D108 insertions in the plasmid pSC101
AU - Szatmari, George B.
AU - Kahn, Jeffrey
AU - DuBow, Michael S.
N1 - Funding Information:
The authors would like to thank Martine Lapointe for her expertt echnical assistance.T his researchw as supported by grants from the Medical Research Council of Canada (MA675 l), and a StrategicG rant (G0907) from the Natural Sciences and Engineering Research Council of Canada. G.B.S. was supported by a postgraduate scholarship from the Natural Sciences and Engineering Research Council of Canada. M.S.D. is a Scholar of the Medical Research Council of Canada.
PY - 1986
Y1 - 1986
N2 - We have isolated four independent insertions of the entire 37-kb D10Scts10 genome in the low-copy-number plasmid pSC101 in vivo. They were all formed by replicative transposition during the D108 lytic cycle. The orientation of these four insertions was found to be the same, with the left ends facing towards pS101 replication, and the right end facing in the direction of all pS101 transcription, as was previously found for a Mucts62 insertion in pS101, pMC321. The exact sites of insertion of two of the D108 prophages, as well as the Mu prophage, have been determined by sequence analysis. All three insertions caused a 5-bp duplication of pS101 sequences at the target site, as has been found for insertions formed by conservative integration upon lysogeny. Moreover, we have determined the nucleotide sequence of the first 75 bp of the right end of D108 and, though this end is interchangeable with the right end of Mu as a substrate for either phage's transposition functions, there are a number of nucleotide differences between them.
AB - We have isolated four independent insertions of the entire 37-kb D10Scts10 genome in the low-copy-number plasmid pSC101 in vivo. They were all formed by replicative transposition during the D108 lytic cycle. The orientation of these four insertions was found to be the same, with the left ends facing towards pS101 replication, and the right end facing in the direction of all pS101 transcription, as was previously found for a Mucts62 insertion in pS101, pMC321. The exact sites of insertion of two of the D108 prophages, as well as the Mu prophage, have been determined by sequence analysis. All three insertions caused a 5-bp duplication of pS101 sequences at the target site, as has been found for insertions formed by conservative integration upon lysogeny. Moreover, we have determined the nucleotide sequence of the first 75 bp of the right end of D108 and, though this end is interchangeable with the right end of Mu as a substrate for either phage's transposition functions, there are a number of nucleotide differences between them.
KW - 5 base pair duplications of target
KW - In vivo DNA cloning
KW - cointegrate transposition
KW - recombinant DNA
KW - restriction mapping
KW - right end sequence
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U2 - 10.1016/0378-1119(86)90113-7
DO - 10.1016/0378-1119(86)90113-7
M3 - Article
C2 - 3011604
AN - SCOPUS:0022521989
SN - 0378-1119
VL - 41
SP - 315
EP - 319
JO - Gene
JF - Gene
IS - 2-3
ER -