Abstract：
Polyacrylamide gel electrophoresis is commonly used to characterize the chain length of polyphosphates (polyP), more generally called condensed phosphates. After separation, nonradioactive, optical polyP staining is limited to chain lengths greater than 15 PO 3 - ${\\rm{PO}}_3^ - $ monomers with toluidine blue or 4\',6-diamidino-2-phenylindole. PolyP chain lengths longer than 62 PO 3 - $\\;{\\rm{PO}}_3^ - $ monomers were correlated to the shortest DNA ladders. In this study, synthetic linear polyPs (Sigma-Aldrich \"Type 45\", estimated mean length of 45 PO 3 - ${\\rm{PO}}_3^ - $ monomers), trimetaphosphate (trimetaP: 3 PO 3 - ${\\rm{PO}}_3^ - $ ring), tripolyphosphate (tripolyP), pyrophosphate (PPi ), and inorganic orthophosphate (o-Pi ) were visualized after separation by an in situ hydrolytic degradation process to o-Pi that was subsequently stained with methyl green. Statistically insignificant migration reduction of synthetic short-chain polyP after perchloric acid or phenol-chloroform extraction was confirmed with the Friedman test. 31 P diffusion-ordered NMR spectroscopy confirmed that extraction also reduced PPi diffusivity by <10%. Linear regression between the Rf peak migration value and the logarithm of synthetic polyP molecular weights enabled estimation of extracted polyP chain lengths from 2 to 45 PO 3 - ${\\rm{PO}}_3^ - $ monomers. Linear polyP extracts from Saccharomyces cerevisiae grown in aerobic conditions were generally shorter than extracts cultured in anaerobic conditions. Extractions from both aerobic and anaerobic S. cerevisiae included tripolyP and o-Pi , but no PPi .
摘要：
聚丙烯酰胺凝胶电泳通常用于表征多磷酸盐（polyP）的链长，通常称为缩合磷酸盐。分离后,非放射性,光学polyP染色限于链长大于15PO3-${\\rm{PO}}_3^-具有甲苯胺蓝或4'的单体，6-二氨基-2-苯基吲哚。大于62PO3-$\\;{\\rm{PO}}_3^-$单体的PolyP链长度与最短的DNA阶梯相关。在这项研究中,合成线性聚合物(Sigma-Aldrich“45型”，估计45PO3-${\\rm{PO}}_3^-$单体的平均长度)，三偏磷酸盐(三偏磷酸盐:3PO3-${\\rm{PO}}_3^-$环),三聚磷酸盐（三聚P），焦磷酸盐(PPi)，和无机正磷酸盐（o-Pi）在通过原位水解降解过程分离为o-Pi后可视化，然后用甲基绿染色。用Friedman试验证实了高氯酸或苯酚-氯仿萃取后合成短链polyP的统计学上微不足道的迁移减少。31P扩散有序NMR光谱证实萃取也将PPi扩散系数降低<10%。Rf峰迁移值与合成聚P分子量的对数之间的线性回归能够估计从2到45PO3-${\\rm{PO}}_3^-$单体的提取聚P链长度。在有氧条件下生长的酿酒酵母的线性polyP提取物通常比在厌氧条件下培养的提取物短。好氧和厌氧酿酒酵母的提取物包括三聚P和o-Pi，但没有PPI。
