TY - JOUR
T1 - Genome locations of temperature-sensitive mutants in glycoprotein gB of herpes simplex virus type 1
AU - DeLuca, Neal
AU - Person, Stanley
AU - Bzik, David J.
AU - Snipes, Wallace
N1 - Funding Information:
This work was supported by the Ernest W. Peters Memorial grant from the American Cancer Society.
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1984/9
Y1 - 1984/9
N2 - A plasmid containing a herpes simplex virus type 1 (HSV-1) insert from strain KOS, prototypic coordinates 0.345 to 0.368 (3.45 kilobases) was mutagenized in vitro, and potential mutations were introduced into intact viral DNA by cotransfection. Functions normally associated with the glycoprotein gB are in the 1-9 complementation group, and the above coordinates include those that specify the gB glycoprotein gene. Following cotransfection, individual plaques were screened for temperature sensitivity (ts) of viral growth. A total of seven ts mutants was obtained, of which four were spurious mutations due to alterations outside the cloned sequences, presumably mediated by some aspect of the Ca-precipitation-cotranafection method. The remaining three did not complement known mutants of the 1-9 complementation group. These three mutants, along with tsJ12 (P. A. Schaffer, G. M. Aron, N. Biswal, and M. Benyesh-Melnick, 1973, Virology 52, 57-71) and tsJ33 (C.-T. Chu, D. S. Parris, R. A. F. Dixon, F. E. Farber, and P. A. Schaffer,1979, Virology 98, 168-181), were physically located by marker-rescue experiments to three different restriction fragments between 0.345 to 0.368 map units. Sodium dodecyl sulfate-gel electrophoresis was used to analyze the glycoproteins synthesized during continuous or pulse-chase labeling protocols. All five mutants were found to synthesize a precursor of gB but did not accumulate mature gB during a pulse, a chase, or continuous labeling at the nonpermissive temperature.
AB - A plasmid containing a herpes simplex virus type 1 (HSV-1) insert from strain KOS, prototypic coordinates 0.345 to 0.368 (3.45 kilobases) was mutagenized in vitro, and potential mutations were introduced into intact viral DNA by cotransfection. Functions normally associated with the glycoprotein gB are in the 1-9 complementation group, and the above coordinates include those that specify the gB glycoprotein gene. Following cotransfection, individual plaques were screened for temperature sensitivity (ts) of viral growth. A total of seven ts mutants was obtained, of which four were spurious mutations due to alterations outside the cloned sequences, presumably mediated by some aspect of the Ca-precipitation-cotranafection method. The remaining three did not complement known mutants of the 1-9 complementation group. These three mutants, along with tsJ12 (P. A. Schaffer, G. M. Aron, N. Biswal, and M. Benyesh-Melnick, 1973, Virology 52, 57-71) and tsJ33 (C.-T. Chu, D. S. Parris, R. A. F. Dixon, F. E. Farber, and P. A. Schaffer,1979, Virology 98, 168-181), were physically located by marker-rescue experiments to three different restriction fragments between 0.345 to 0.368 map units. Sodium dodecyl sulfate-gel electrophoresis was used to analyze the glycoproteins synthesized during continuous or pulse-chase labeling protocols. All five mutants were found to synthesize a precursor of gB but did not accumulate mature gB during a pulse, a chase, or continuous labeling at the nonpermissive temperature.
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U2 - 10.1016/0042-6822(84)90230-7
DO - 10.1016/0042-6822(84)90230-7
M3 - Article
C2 - 6091335
AN - SCOPUS:0021127828
SN - 0042-6822
VL - 137
SP - 382
EP - 389
JO - Virology
JF - Virology
IS - 2
ER -