Kenny-Ganzert Isabel, Chi Qiuyi, Sherwood David
Department of Biology, Duke University.
MicroPubl Biol. 2023 Mar 21;2023. doi: 10.17912/micropub.biology.000813. eCollection 2023.
Transgene driven protein expression is an important tool for investigating developmental mechanisms in . Here, we have assessed protein production rates and levels in L3 larval uterine cells (UCs). Using ubiquitous promoter driven cytosolic and transmembrane tethered GFP, fluorescence recovery after photobleaching, and quantitative fluorescence analysis, we reveal that cytosolic GFP is produced at an ~two-fold higher rate than transmembrane tethered GFP and accumulates at ~five-fold higher levels in UCs. We also provide evidence that cytosolic GFP in the anchor cell, a specialized UC that mediates uterine-vulval connection, is more rapidly degraded through an autophagy-independent mechanism.
转基因驱动的蛋白质表达是研究发育机制的重要工具。在此，我们评估了L3幼虫子宫细胞（UCs）中的蛋白质产生速率和水平。通过使用普遍启动子驱动的胞质和跨膜连接的绿色荧光蛋白（GFP）、光漂白后的荧光恢复以及定量荧光分析，我们发现胞质GFP的产生速率比跨膜连接的GFP高约两倍，并且在UCs中的积累水平高约五倍。我们还提供证据表明，在介导子宫 - 外阴连接的特殊UC——锚定细胞中的胞质GFP，通过一种不依赖自噬的机制被更快地降解。
