TY - JOUR
T1 - Transposition of λplacMu is mediated by the A protein altered at its carboxy-terminal end
AU - Bremer, Erhard
AU - Silhavy, Thomas J.
AU - Weinstock, George M.
N1 - Funding Information:
We thank M. Berman, M. Howe and R. Zagursky for providing strains and bacteriophages and S. Beck for advice on DNA sequencing.W e are grateful to D. Kamp for the communication of data on the Mu4 sequence prior to publication. We thank M. Manson and G. Sweet for critical reading of the manuscript, V. Koogle for help in preparing the manuscript, and A. Middendorf for technical assistance. E.B. thanks W. Boos for his support. This work was supported in part by the National Cancer Institute, Department of Health and Human Services, under Contract No. NOl-CO-23909 with Litton Bionetics, and in part by the Deutsche Forschungsgemeinschaft through SFB 156. E.B. was the recipient of a fellowship from the Deutsche Akademische Austauschdienst.
PY - 1988/11/15
Y1 - 1988/11/15
N2 - λplacMu phages are derivatives of bacteriophage λ that use the transposition machinery of phage Mu to insert into chromosomal and cloned genes. When inserted in the proper fashion, these phages yield stable fusions to the Escherichia coli lac operon in a single step. We have determined the amount of DNA from the c end of phage Mu present in one of these phages, λplacMu3, and have shown that this phage carries a 3137-bp fragment of Mu DNA. This DNA segment carries the Mu c-end attachment site and encodes the Mu genes cts62, ner+, and gene A lacking 179 bp at its 3′ end (A′). The product of this truncated gene A′ retains transposase activity and is sufficient for the transposition of λplacMu. This was demonstrated by showing that λplacMu derivatives carrying the A am1093 mutation in the A′ gene are unable to transpose by themselves in a Su- strain, but their transposition can be triggered by coinfection with λpMu507(A+ B+). We have constructed several new λplacMu phages that carry the A′ am1093 gene and the kan gene, which confers resistance to kanamycin. Chromosomal insertions of these new phages are even more stable than those of the previously reported λplacMu phages, which makes them useful tools for genetic analysis.
AB - λplacMu phages are derivatives of bacteriophage λ that use the transposition machinery of phage Mu to insert into chromosomal and cloned genes. When inserted in the proper fashion, these phages yield stable fusions to the Escherichia coli lac operon in a single step. We have determined the amount of DNA from the c end of phage Mu present in one of these phages, λplacMu3, and have shown that this phage carries a 3137-bp fragment of Mu DNA. This DNA segment carries the Mu c-end attachment site and encodes the Mu genes cts62, ner+, and gene A lacking 179 bp at its 3′ end (A′). The product of this truncated gene A′ retains transposase activity and is sufficient for the transposition of λplacMu. This was demonstrated by showing that λplacMu derivatives carrying the A am1093 mutation in the A′ gene are unable to transpose by themselves in a Su- strain, but their transposition can be triggered by coinfection with λpMu507(A+ B+). We have constructed several new λplacMu phages that carry the A′ am1093 gene and the kan gene, which confers resistance to kanamycin. Chromosomal insertions of these new phages are even more stable than those of the previously reported λplacMu phages, which makes them useful tools for genetic analysis.
KW - Mu transposase
KW - bacteriophages λ and Mu
KW - lacZ fusions
KW - recombinant DNA
KW - transducing phages
UR - http://www.scopus.com/inward/record.url?scp=0024206792&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0024206792&partnerID=8YFLogxK
U2 - 10.1016/0378-1119(88)90089-3
DO - 10.1016/0378-1119(88)90089-3
M3 - Article
C2 - 2850974
AN - SCOPUS:0024206792
SN - 0378-1119
VL - 71
SP - 177
EP - 186
JO - Gene
JF - Gene
IS - 1
ER -