DeLuca N, Person S, Bzik D J, Snipes W
Virology. 1984 Sep;137(2):382-9. doi: 10.1016/0042-6822(84)90230-7.
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-cotransfection 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.
一个含有来自KOS株单纯疱疹病毒1型（HSV-1）插入片段（原型坐标0.345至0.368，3.45千碱基）的质粒在体外被诱变，并通过共转染将潜在突变引入完整的病毒DNA中。通常与糖蛋白gB相关的功能属于1-9互补组，上述坐标包括指定gB糖蛋白基因的那些坐标。共转染后，筛选各个噬斑的病毒生长温度敏感性（ts）。总共获得了7个ts突变体，其中4个是由于克隆序列外的改变导致的假突变，推测是由Ca沉淀-共转染方法的某些方面介导的。其余3个不与1-9互补组的已知突变体互补。这3个突变体，连同tsJ12（P.A. Schaffer、G.M. Aron、N. Biswal和M. Benyesh-Melnick，1973年，《病毒学》52卷，57-71页）和tsJ33（C.-T. Chu、D.S. Parris、R.A.F. Dixon、F.E. Farber和P.A. Schaffer，1979年，《病毒学》98卷，168-181页），通过标记拯救实验在0.345至0.368图谱单位之间的三个不同限制片段上进行了物理定位。使用十二烷基硫酸钠-凝胶电泳分析在连续或脉冲追踪标记方案期间合成的糖蛋白。发现所有5个突变体都合成了gB的前体，但在非允许温度下的脉冲、追踪或连续标记过程中没有积累成熟的gB。
