Mojica-A T, Middleton R B
J Bacteriol. 1971 Dec;108(3):1161-7. doi: 10.1128/jb.108.3.1161-1167.1971.
At least one factor that causes low fertility of Salmonella typhimurium LT2 strains in crosses with Escherichia coli K-12 Hfr's can be inhibited by growing the female strains in supplemented minimal salts medium rather than in nutrient broth and by incubating the female strains at 50 C immediately before mating with the Hfr. These pretreatments can enhance the recovery of prototrophic recombinants for markers injected early by the Hfr by a factor of as much as 10(4). The heat treatment is effective only on the female in intergeneric crosses and gradually loses (within 50 min) its effectiveness after return of heat-treated cells to 37 C. It is concluded that the restriction system of the female is heat-sensitive. Since markers injected late by the male enter females in which the heat-impaired restriction system has recovered, few recombinants for late markers are found. The presence of the leading end of an E. coli Hfr in an S. typhimurium-E. coli hybrid enhances by up to sevenfold the frequency of lac(+) recombinants in subsequent crosses with an E. coli Hfr if the E. coli segment is integrated into the chromosome of the hybrid; the effect is less marked if the E. coli segment is not integrated.
鼠伤寒沙门氏菌LT2菌株与大肠杆菌K - 12 Hfr杂交时导致育性低的至少一个因素,可以通过将雌性菌株在添加了营养成分的基本盐培养基中培养,而不是在营养肉汤中培养,并在与Hfr交配前立即将雌性菌株在50℃下孵育来抑制。这些预处理可以使Hfr早期注入的标记的原养型重组体的回收率提高多达10⁴倍。热处理仅对属间杂交中的雌性有效,并且在热处理后的细胞回到37℃后(50分钟内)逐渐失去其有效性。可以得出结论,雌性的限制系统对热敏感。由于雄性后期注入的标记进入热损伤的限制系统已经恢复的雌性中,所以很少发现后期标记的重组体。如果大肠杆菌片段整合到杂交体的染色体中,在鼠伤寒沙门氏菌 - 大肠杆菌杂交体中存在大肠杆菌Hfr的前端会使随后与大肠杆菌Hfr杂交时lac⁺重组体的频率提高多达七倍;如果大肠杆菌片段未整合,这种效果则不太明显。