AI Article Synopsis
- Drosophila retrotransposon copia produces virus-like particles (VLPs) essential for its replication, containing copia RNA and reverse transcriptase.
- A new, efficient method for purifying these VLPs from Drosophila S2 cells is introduced, streamlining the process by using simple centrifugation instead of complex gradients.
- The study confirms the presence and purity of VLPs through multiple analysis techniques and reveals that copia particles are primarily found in the nucleus of the cells.
Article Abstract
Drosophila retrotransposon copia produces virus-like particles (VLPs) in cultured Drosophila cells. The VLPs contain copia RNA and reverse transcriptase activity, and thus, play a major role in copia replication. Here, we report a rapid and simple method for the purification of copia VLPs from Drosophila Schneider 2 (S2) cells. The VLP purification procedure consists of a series of short centrifugation steps and eliminates the tedious and time-consuming performance of sucrose, metrizamide and cesium chloride (CsCl) gradients. The purity and presence of VLPs at different purification steps was monitored by transmission electron microscopy (TEM), immunochemical detection methods, and by Northern blot analysis. In addition to providing a fast protocol for VLP purification, our results also show that copia particles are mainly located in the nucleus of S2 cells. This new protocol may find broad applications for the purification of various other VLPs, and thus, may be of great value to other investigators with similar interests.
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| http://dx.doi.org/10.1016/j.jviromet.2003.09.025 | DOI Listing |
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