Gustafsson P, Nordström K, Normark S
J Bacteriol. 1973 Nov;116(2):893-900. doi: 10.1128/jb.116.2.893-900.1973.
Wild-type strains of Escherichia coli K-12 adsorb gentian violet to the cell surface, but the dye is not transported into the cytoplasm. However, when some mutants that have an altered outer membrane are exposed to gentian violet, the dye is also found in the ribosomal fraction. The transport into the cytoplasm is inhibited at 0 C and requires that the concentration of gentian violet exceeds a threshold value. The initial rate of uptake as well as the amount of gentian violet found in the cytoplasm increases with the concentration of the dye in the medium. The rate of transport of the dye into the cytoplasm is much lower for stationary mutant cells than for exponentially growing cells. The rate of uptake into the cytoplasm increases with increasing deficiency of carbohydrate in the lipopolysaccharide (carbohydrate content lpsB > lpsA > galU). However, other components are also responsible for the barrier since an envA mutant which is not altered in the lipopolysaccharide carbohydrates show an extremely rapid uptake of the dye. The rate of uptake for the envA mutant was the highest found and the same as that of spheroplasts. Growth in the presence of agents affecting the murein sacculus, e.g., lysozyme and sublethal concentrations of penicillin, increased the rate of uptake of gentian violet. Brief treatments with tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetic acid drastically impaired the barrier function. Inhibition of protein synthesis by chloramphenicol also opened the barrier to gentian violet. In conclusion, the outer part of the bacterial envelope is a penetration barrier for gentian violet and probably also for other substances. The lipopolysaccharide, the murein and also other components are important for the function of this barrier. Resistance to gentian violet was found to be inversely correlated to the rate of penetration of the dye into the cytoplasm.
大肠杆菌K - 12的野生型菌株会将结晶紫吸附到细胞表面，但这种染料不会转运到细胞质中。然而，当一些外膜发生改变的突变体暴露于结晶紫时，在核糖体部分也能发现这种染料。转运到细胞质中的过程在0℃时受到抑制，并且需要结晶紫的浓度超过阈值。初始摄取速率以及在细胞质中发现的结晶紫量会随着培养基中染料浓度的增加而增加。对于静止的突变细胞，染料转运到细胞质中的速率远低于指数生长的细胞。随着脂多糖中碳水化合物缺乏程度的增加（碳水化合物含量lpsB > lpsA > galU），摄取到细胞质中的速率也会增加。然而，其他成分也对这种屏障起作用，因为在脂多糖碳水化合物方面未发生改变的envA突变体对染料的摄取极其迅速。envA突变体的摄取速率是所发现的最高速率，与原生质体的摄取速率相同。在存在影响胞壁质囊的试剂（如溶菌酶和亚致死浓度的青霉素）的情况下生长，会增加结晶紫的摄取速率。用三（羟甲基）氨基甲烷 - 乙二胺四乙酸进行短暂处理会严重损害屏障功能。氯霉素抑制蛋白质合成也会打开对结晶紫的屏障。总之，细菌包膜的外部是结晶紫以及可能对其他物质的渗透屏障。脂多糖、胞壁质以及其他成分对该屏障的功能都很重要。发现对结晶紫的抗性与染料渗透到细胞质中的速率呈负相关。
