人類的生命週期中，類固醇賀爾蒙決定一個個體發育至各種特定階段，然而其分子機制仍然未明確，為了了解類固醇賀爾蒙如何調節發育，我們使用果蠅的邊境細胞作為實驗模型進行研究，這些細胞會因蛻皮激素的調控進行遷移。除此之外，透過僅表現同型合子droj2的突變在邊境細胞的實驗中，也確認只有droj2的突變情況下，也會導致邊境細胞的移動缺陷。關於droj2，它是人類熱休克蛋白40 （HSP40）的同源基因，其功能是為避免不正常摺疊蛋白質的聚集以及將這些不正常的折疊蛋白恢復成適合的結構，而其他系統的HSP40也負責賀爾蒙受器的成熟。雖然果蠅中DROJ2與類固醇賀爾蒙受器的關係還不是很清楚，但這些研究，也許暗示了DROJ2與蛻皮激素受體是有交互關係，為了驗證這個假說，利用蛻皮激素反應元件所調控的lacZ （EcRElacZ）基因表現作為報告者基因，發現droj2的突變會在Tai（△B）大量表現下，使EcRElacZ的表現也會加強。而我們也透過建構重組蛋白GST-DROJ2以及蛻皮激素受體:His-tag-EcR.A、His-tag-EcR.B1、His-tag-EcR.A△HBD、His-tag-EcR.B1△HBD以及His-tag-EcR△N，透過GST-pull down assay，來了解DROJ2跟蛻皮激素在生化上的交互關係。在未來透過這樣的研究知道DROJ2跟蛻皮激素受體是如何調節蛻皮激素訊息的傳遞，將會提供去理解卵子生成又或者是動物發育的過程。

DROJ2 is a heat shock protein 40 （HSP40）in Drosophila. DROJ2 function is critical in protein stability, cell physiology including oogenesis and cell migration. However, the mechanism is unclear for DROJ2 for collective cell migration. Previous reports show that HSP40 control the stability of steroid hormone receptor. Interestingly, the detachment timing of border cell migration is regulated by ecdysone, the only known steroid hormone in Drosophila. In previous work, we reported that homozygous mutant clones of droj2 caused the border cell migration defect. Thus, we hypothesized that DROJ2 regulates border cell migration via control the stability of ecdysone receptor to modulate the ecdysone signaling level. To assess this possibility, EcRElacZ, a transgenic reporter of ecdysone reporter, was applied to assess the effect of droj2. The results indicate that EcRElacZ was increased under the droj2GS5242 genetic background. To explore the physical interaction between the ecdysone receptor and DROJ2, the recombinant protein, GST-DROJ2, was generated to perform GST pull down assay. In addition, full-length of ecdysone receptors, His-tag EcR.A and His-tag EcR.B1, were also constructed for protein/protein interaction. I also constructed three truncated forms of the ecdysone receptor, His-tag EcR.A△HBD, His-tag EcR.B1△HBD, and His-tag EcR△N, for analyzing domain requirement for EcRA and DROJ2. Detail analysis will be presented in main results.

余惠穎，分析果蠅的群體細胞遷移中DROJ2的功能，國立成功大學生物科技與產業科學系碩士論文，2018。
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