In vitro transcription assay for the human placenta-specific transcription factor, hGCMa.

• Main Authors: Chuancheng Lin, 林泉成
• Other Authors: Hungwen Chen
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/21708431676625810532

碩士 === 國立臺灣大學 === 生化科學研究所 === 91 === GCMa belongs to a novel family of zinc-containing transcription factors and is a placenta-specific transcription factor. GCMa primarily expresses in placental trophoblasts and is involved in placental development. Syncytin is the envelope protein of the human endogenous retrovirus family W (HERV-W). Syncytin primarily expresses in the syncytiotrophoblast layer and mediates fusion of cytotrophoblasts into the syncytiotrophoblast layer. Previously, we have demonstrated that hGCMa regulates syncytin gene expression and the syncytin-mediated trophoblastic fusion. To further understand the molecular mechanism of hGCMa transcriptional activation, in this thesis, an in vitro transcription assay for hGCMa was successfully established using G-free cassette reporter plasmids containing multi-copies of hGCMa-binding site, HeLa nuclear extracts, and recombinant hGCMa protein. In addition, we demonstrated that recombinant hGCMa protein prepared from the baculovirus-insect cell and E. coli expression systems is biological active in terms of transcriptional activity. The established in vitro transcription assay may help future study on the regulation of hGCMa transcriptional activity.