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Fig. 5. Sedimentation analysis of GFP-SRP19 present in cytoplasm isolated from injected oocytes. The injected oocytes were either immediately enucleated and frozen or incubated for 48 hours before enucleation and freezing. Half of the 48 hour sample was treated with ribonuclease A (RNase) for 30 minutes in buffer lacking Mg²⁺. Clarified cytoplasms from 25 oocytes taken immediately after injection (open circles), after 48 hours (filled circles) and after 48 hours and treated with RNase (open squares) were loaded on glycerol gradients and centifuged under the same conditions as described in Fig. 1. Gradient fractions were collected and (A) scanned at for fluorescence at 510 nm (FL, arbitrary units). Fractions from parallel gradients loaded with the sedimentation markers described in Fig. 1 were scanned at 280 nm (M_r, open triangles). The predicted sedimentation position of SRP (11 S, M_r 335,000) is indicated by an arrow. Fractions were then precipitated with acetone and (B) analysed by immunoblotting using antibodies to GFP or, in the case of markers, staining of the gel with Coomassie brilliant blue.

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