White L A, MacLeod R A
Appl Microbiol. 1971 Mar;21(3):520-6. doi: 10.1128/am.21.3.520-526.1971.
The uptake of phosphate, from media limited in this ion, by resting cells of Aerobacter aerogenes has been investigated and shown to be dependent upon several factors. An incubation medium composed of 10(-2)m K(+), 5 x 10(-3)m Mg(2+), 1 mg of glucose per ml, and 1 muCi of (32)PO(4) (3-) per ml, buffered at pH 6.55 with 0.05 mN-2-hydroxyethyl-piperazine-N'-2-ethanesulfonic acid (HEPES), was found to stimulate optimum accumulation of (32)P-orthophosphate. The temperature of incubation, incubation time, the concentration of unlabeled orthophosphate, as well as arsenate, and several metabolic inhibitors were found to affect the accumulation. The labeled cells were collected on a membrane filter, which had been previously boiled in glass-distilled water, for measurement of the radioactivity accumulated. Under optimum conditions, as few as 20,000 cells were capable of accumulating detectable amounts of (32)P-orthophosphate in 1 hr of incubation.
已对产气气杆菌静息细胞从该离子受限的培养基中摄取磷酸盐的情况进行了研究，结果表明其摄取取决于几个因素。发现一种由10⁻²m K⁺、5×10⁻³m Mg²⁺、每毫升1毫克葡萄糖和每毫升1微居里³²PO₄³⁻组成的孵育培养基，用0.05 mN - 2 - 羟乙基哌嗪 - N' - 2 - 乙烷磺酸（HEPES）在pH 6.55缓冲，可刺激³²P - 正磷酸盐的最佳积累。发现孵育温度、孵育时间、未标记正磷酸盐以及砷酸盐的浓度，还有几种代谢抑制剂都会影响积累。将标记的细胞收集在预先在玻璃蒸馏水中煮沸过的膜滤器上，用于测量积累的放射性。在最佳条件下，孵育1小时，低至20000个细胞就能积累可检测量的³²P - 正磷酸盐。
