Jensen J L, Susman M
Genetics. 1980 Feb;94(2):301-25. doi: 10.1093/genetics/94.2.301.
After nitrosoguanidine mutagenesis, a Phage Host Defective (phd) mutant of E. coli HfrH was isolated that supported the growth of T4D wild-type bacteriophage at 30 degrees, but not at 40 degrees or higher. Eleven independent spontaneous mutants of T4 (go mutants) were isolated that overcame the growth restriction at high temperature. All of these mutants were located within three percent recombination of a gene 39 amber mutation in the clockwise direction on the standard map. In mixed infections, the representative go mutant chosen for further study seems to be recessive to its wild-type allele. Temperature-shift experiments suggested that the mutated host function involved in phage growth is a "late" function, beginning in mid-eclipse.--Electrophoresis of phage proteins labelled early and late in infection showed that under restrictive conditions early protein synthesis was normal, but that certain late proteins were absent. However, measurements of DNA synthesis showed that under restrictive conditions the amount of phage DNA synthesized, and especially the amount of DNA sedimenting as high molecular weight replicative intermediate, was reduced. Pulse-chase experiments showed that the phage DNA made under restrictive conditions was not rapidly degraded.
经亚硝基胍诱变后,分离出大肠杆菌HfrH的噬菌体宿主缺陷型(phd)突变体,该突变体在30摄氏度时能支持T4D野生型噬菌体生长,但在40摄氏度或更高温度下则不能。分离出11个独立的T4自发突变体(go突变体),它们克服了高温下的生长限制。所有这些突变体在标准图谱上位于基因39琥珀突变顺时针方向3%重组范围内。在混合感染中,选择用于进一步研究的代表性go突变体似乎对其野生型等位基因呈隐性。温度转换实验表明,参与噬菌体生长的突变宿主功能是一种“晚期”功能,始于感染中期。——感染早期和晚期标记的噬菌体蛋白质电泳显示,在限制条件下,早期蛋白质合成正常,但某些晚期蛋白质缺失。然而,DNA合成测量表明,在限制条件下,合成的噬菌体DNA量,尤其是作为高分子量复制中间体沉降的DNA量减少。脉冲追踪实验表明,在限制条件下产生的噬菌体DNA不会迅速降解。