. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 1 2 3 4567 Fraction 8 9 Figure 1. Hydrophobic-interaction chromatography. The clarified demineralized nitrate (4 1) was adjusted to 2 M NaCl, adsorbed to 8 g of the t-butyl HIC resin, poured into a column, and eluted with Tris Cl buffer as described in Materials and Methods. Fractions of 12 ml were collected; ml of each fraction was assayed for morphogenic activity (solid line). Absorbance at 212 nm is shown (dashed line).. Morphogen (ml) Figure 2. Activity of the HIC-punhed and concentrated morphogen. Sample
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 1 2 3 4567 Fraction 8 9 Figure 1. Hydrophobic-interaction chromatography. The clarified demineralized nitrate (4 1) was adjusted to 2 M NaCl, adsorbed to 8 g of the t-butyl HIC resin, poured into a column, and eluted with Tris Cl buffer as described in Materials and Methods. Fractions of 12 ml were collected; ml of each fraction was assayed for morphogenic activity (solid line). Absorbance at 212 nm is shown (dashed line).. Morphogen (ml) Figure 2. Activity of the HIC-punhed and concentrated morphogen. Samples ( mil of the pooled morphogen eluted in the peak frac- tions from the HIC resin (fractions 2-5 from Fig. I) were assayed in duplicate for morphogenic activity. Controls = ml Tris Cl buffer with no morphogen. Error bars indicate deviation from the mean. The yield of morphogen that can be solubilized (and subsequently purified by HIC) is strongly dependent on the concentration of chelator in the initial extract (Table I). The results shown in Table IA demonstrate that rela- tively little inducer is solubilized by prolonged (20 h) stir- ring of the water-washed crude cell wall fraction in aqueous buffer in the absence of chelator, and that solu- bilization increases markedly when chelator ( M EDTA) is present. The absolute concentration of the che- lator also is critical to obtain high yields of the soluble morphogen. Thus (Table IB), in three parallel extractions using the same total amount of chelator (50 mmole EDTA/g alga), sufficient to completely decalcify the cell walls in all three samples, progressively lower yields of soluble inducer are obtained with progressively higher concentrations of EDTA. This behavior is consistent with the hydrophobic property of the amphipathic morphogen; at progressively higher concentrations of the highly charged EDTA salt [nominally, (Na+)4 EDTA 4], the hy- drophobic behavior of the morphogen becomes increas- ingly dominant,
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
