| Autor: |
Cervera, Amelia, Denisse Urbina, Peña, Marcos De La |
| Rok vydání: |
2016 |
| DOI: |
10.6084/m9.figshare.c.3612908_d6.v1 |
| Popis: |
Circular and linear retrozyme RNAs in J. curcas and F. ananassa. A: Double PAGE analysis of an RNA extract (~20 μg) from J. curcas leaves. A gel stripe from a 5 % native polyacrylamide gel containing RNAs of 600–900 nt (left) was cut out and run on top of a second 5 % denaturing polyacrylamide gel (center). The corresponding Northern blot (right), using a digoxigenin-labelled J. curcas retrozyme fragment as a probe, revealed the presence of both circular and linear RNA forms. B: Northern blot analysis of RNA extracts (~30 μg each) from J. curcas leaves, young seedlings and seeds. Samples were run on a 5 % denaturing PAGE and were detected using the same probe as in panel A. C: An in vitro transcription of a previously cloned full genomic retrozyme of F. ananassa was run on a 5 % denaturing PAGE. The RNAs corresponding to the uncleaved full RNA (1134 nt) and double self-cleaved retrozyme RNA (679 nt) were cut out and purified (marked in red). D: Purified retrozyme RNA (679 nt) was circularized with a tRNA ligase for 2 h and run on a 5 % denaturing PAGE. The circularized RNA (upper band) was cut out and purified. E: Northern blot hybridization of a F. ananassa RNA extract (~30 μg) and previously purified RNA markers (linear, circular and uncleaved full retrozyme RNAs, ~1 ng each) run on a 5 % native PAGE. F: Northern blot hybridization of the same RNA samples as in panel E run on a 5 % denaturing PAGE. Ethidium bromide staining of the 5S rRNA is shown at the bottom as a loading control. (PDF 728 kb) |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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