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Thesis videos

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posted on 27.05.2022, 13:44 authored by Daisy VinterDaisy Vinter

  

Movie 1: Maximum intensity projection of a representative embryo showing hb translation sites (grey) and Histone-RFP (magenta) through nc11-14. Embryos are from females with single copies of scFv-mNG-NLS and His-RFP crossed to males with the hbP2>SunTag-hb transgene, and are imaged with a 40x objective, 1.1x optical zoom and 46 sec time resolution between frames. Time stamp is in min:sec. 


Movie 2: Single plane time-lapse experiment visualising plasma membrane tethered MCP-mRuby3-CAAX and bound hbP2>SunTag-hb mRNAs (magenta) and translation sites or mature proteins bound by scFv-mNG-NLS (green). Embryos are from females carrying single copies of the scFv-NG and tdMCP-Ruby3-CAAX transgenes crossed to SunTag-hb-MS2 transgenic males. Imaged with a 63x objective, 8x optical zoom, and a time resolution of ~2 sec per frame. Time stamp is in min:sec.  


Movie 3: Single plane time-lapse experiment showing a single protein being released from a translation site (green, scFv-mNG-NLS) and diffusing away from the mRNA (magenta, tdMCP-mRuby3-CAAX) and site of translation. The embryo genotype and imaging settings are as described for movie 3, except the time resolution is ~1.7 sec per frame. 

  

Movie 4: Single plane time-lapse experiment visualising translation termination. An active translation site site (green, scFv-NG) terminates translation and a single protein diffuses away from the mRNA molecule (magenta, MCP-mRuby3-CAAX). The embryo genotype and imaging settings are as for Movie 2.


Movie 5: Enlarged view of Movie 2, highlighting independent translation sites (green) fusing and separating over time. 


Movie 6: Maximum intensity projection of another representative embryo showing hb translation sites (grey) and Histone-RFP (magenta) through nc11-14. Embryos are from females with single copies of scFv-mNG-NLS and His-RFP crossed to males with the hbP2>SunTag-hb transgene, and are imaged with a 40x objective, 1.1x optical zoom and 46 sec time resolution between frames. Time stamp is in min:sec.  


Movie 7: Maximum intensity projection of a representative embryo showing hb translation sites (green) and Histone-RFP (magenta) through nc11-14. Embryos are from females with single copies of scFv-mNG-NLS and His-RFP crossed to males with the hbP2>SunTag-hb-SV40 transgene, and are imaged with a 40x objective, 1.1x optical zoom and 46 sec time resolution between frames. Time stamp is in min:sec.   


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