. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 178 D. E. MORSE ET AL — 345678 Fraction Figure 3. Anion-exchange chromalography. Five ml of the pooled morphogen eluted in the peak fractions from the HIC resin (fractions 2-5 from Fig. 1) was diluted and chromatographed on Trisacryl-DEAE. Elution from the DEAE was performed with a gradient of increasing NaCI concentration (inset) as described in Materials and Methods. Frac- tions of 5 ml were collected; ml of each fraction was assayed for morphogemc activity (solid line). Ahsorbance at 212 nm is shown (dashed line
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 178 D. E. MORSE ET AL — 345678 Fraction Figure 3. Anion-exchange chromalography. Five ml of the pooled morphogen eluted in the peak fractions from the HIC resin (fractions 2-5 from Fig. 1) was diluted and chromatographed on Trisacryl-DEAE. Elution from the DEAE was performed with a gradient of increasing NaCI concentration (inset) as described in Materials and Methods. Frac- tions of 5 ml were collected; ml of each fraction was assayed for morphogemc activity (solid line). Ahsorbance at 212 nm is shown (dashed line); the bulk of the applied absorbance remained bound on the column. behavior is comparable to that of the purified inducer in solution. Morphogen-based larval flypaper To produce a practical experimental tool that could be deployed in the natural ocean environment, we incor- Table II The purified morphogen is insensitive l irypsin Trypsin treatment Metamorphosis (%) Control Treated 90 ± 10 80 ± 10 HIC-punned morphogen was incubated with immobilized trypsin. separated from the enzyme, and assayed for remaining activity as de- scribed in Materials and Methods. Control was a sample of the untreated morphogen. incubated in parallel, and then combined with a sample of buffer that had been incubated in parallel with the immobilized enzyme. (able 111 Cliclattir xnluhilizcx the parent morplmgen Morphogen Solubilized By Property Chelator1 Enzymes or alkalr M (Da)3 6000-8000 <2000 Stability Higher Low Binding Hydrophobic Yes No Anionic Yes Yes 1 This study. : Data from Morse and Morse, 1991. ' Apparent molecular weight. porated the resin-bound morphogen on a simple, stable platform. We found that the HIC-resin-bound morphogen (prepared from the chromatographically purified mor- phogen) can be attached to surfaces with a waterproof silicone adhesive, without significant loss of activity. The 100 -. Please note that these images are extracted from scanned page images that may have b
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