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Fig. 1. Expression of endogenous SRP RNA and SRP proteins in Xenopus oocytes. (A) Staged oocytes were collected to give an equal volume of soluble extract and RNA was extracted for RT-PCR and quantitation on 2% agarose gels. RT-PCR products were generated using primers specific for SRP RNA (20 cycles) and SRP54 mRNA (25 cycles), and 5S RNA was separated directly from the total RNA sample. (B) Sedimentation analysis of SRP19 protein present in cytoplasms isolated from stage IV oocytes. Gradient fractions were immunoblotted using a chicken antibody directed against SRP19 protein. Reactive bands were detected in fractions sedimenting at 10-12 S. Positions of sedimentation mass markers run in a parallel gradient are indicated by stars (from left: haemoglobin, Mr 
67,000; alcohol dehydrogenase, Mr 180,000; apoferritin, Mr 443,000; microglobulin, Mr 670,000; immunoglobulin M, Mr 960,000: see also Fig. 5). Positions of electrophoretic mass markers are shown (kDa). (C) Immunoblot using anti-SRP19 of the 10-12 S fraction isolated from extracts equivalent to the range of oocyte stages and oocyte numbers described in (A). (D) Relative amounts of product per oocyte were calculated by dividing amounts detected from equal masses of oocytes by the number of oocytes contained in that mass at each stage. Graph shows: SRP RNA (open circles), SRP54 mRNA (open squares), 5 S RNA (black circles) and SRP19 protein (black squares). Intensity of staining with ethidium bromide or recorded on film after ECL was captured using a GeneSnap system and measured using GeneTools (Synoptics Ltd).
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