TY - JOUR
T1 - Identification and restriction endonuclease mapping of the threonine operon regulatory region
AU - Gardner, Jeffrey F.
AU - Reznikoff, William S.
N1 - Funding Information:
%‘e thank 0. Andresson, D. Berg, K. Betrand, R. Blakesley, R. Burgess, S. Hardies, R. Jorgensen. J. Salstrom, and R. Weisberg for helpful discussions, strains, and various enzyme preparations. This work was supported in part by a grant (ROI-GM19670) and a Career Development Award from t,he National Institutes of Health (to W. S. R.). One of ILS (.J. E. tX.) was supported by post-doctoral fellowships from the National Institutes of Health a,nd the American Cancer Society.
PY - 1978/12/5
Y1 - 1978/12/5
N2 - A DNA fragment carrying the thrP region of Escherichia coli has been identified and characterized. Starting from a secondary site lysogen of bacteriophage λ, where the position of the prophage is either within thrP or between thrP and thrP structural genes (Gardner et al., 1974; Gardner & Smith, 1975) it has been possible to isolate transducing phages which carry bacterial DNA from either side of the prophage. By Int-Xis mediated site-specific recombination we have generated recombinant transducing phages which carry an intact thrP region. A phage carrying a regulatory mutation, which maps in the thrP region, has also been constructed. Mapping with several restriction endonucleases has allowed us to construct a map of the cleavage sites of the thrP region. A 1700 base-pair HaeIII fragment carrying the secondary attachment site (attΔOΔ′) was isolated and its position was localized within a 180 base-pair TaqI-HhaI restriction fragment. The location of attΔOΔ′ in the HaeIII fragment suggests that the entire thrP region is carried by this fragment.
AB - A DNA fragment carrying the thrP region of Escherichia coli has been identified and characterized. Starting from a secondary site lysogen of bacteriophage λ, where the position of the prophage is either within thrP or between thrP and thrP structural genes (Gardner et al., 1974; Gardner & Smith, 1975) it has been possible to isolate transducing phages which carry bacterial DNA from either side of the prophage. By Int-Xis mediated site-specific recombination we have generated recombinant transducing phages which carry an intact thrP region. A phage carrying a regulatory mutation, which maps in the thrP region, has also been constructed. Mapping with several restriction endonucleases has allowed us to construct a map of the cleavage sites of the thrP region. A 1700 base-pair HaeIII fragment carrying the secondary attachment site (attΔOΔ′) was isolated and its position was localized within a 180 base-pair TaqI-HhaI restriction fragment. The location of attΔOΔ′ in the HaeIII fragment suggests that the entire thrP region is carried by this fragment.
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U2 - 10.1016/0022-2836(78)90361-3
DO - 10.1016/0022-2836(78)90361-3
M3 - Article
C2 - 368345
AN - SCOPUS:0018221779
SN - 0022-2836
VL - 126
SP - 241
EP - 258
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 2
ER -