肝癌衍生生長因子(hepatoma derived growth factor;HDGF)是一種新的生長因子，於肝癌細胞株中被發現，具有刺激纖維母細胞和肝癌細胞生長的能力。HDGF在結構、氨基酸序列和功能上類似於HMG蛋白。HDGF被認為在細胞週期和腫瘤增殖上扮演關鍵的角色。最近更發現HDGF在腎臟胚胎發育中扮演重要角色，並認為此因子為血管內皮增生因子。此外HDGF與細胞間的相互作用(cell-cell interaction)及細胞移動(cell migration)有關；除此之外HDGF在血管的發育及疾病發生時具有刺激平滑肌細胞增殖的能力，且HDGF帶有兩個細胞核定位訊號(nuclear localization sequence；NLS)序列具nuclear targeting之功能，可刺激細胞生長。 本研究中以E. coli BL21(DE3) 作為表達重組蛋白HDGF的菌株，從不同發酵溫度的改變中及透過不同發酵容器過程中成本之計算，尋找出表達重組蛋白的最佳製程。

HDGF (hepatoma-derived growth factor) is a novel growth factor,identified from conditioned medium of hepatoma cell line. HDGF has growth stimulating activity for fibroblast and some hepatoma cells. HDGF, a novel defined growth factor with mitogenic effect, has homology protein sequence as HMG (high
mobility group) protein and their three dimension structures appeared to be similar to each other. Recently, elevated HDGF expression was found in developing kidneys but less was found in adult kidney. In addition, HDGF expression was found to be
correlated with angiogenic status of tissues. Thus, it is
speculated that HDGF plays a role during embryonic development and angiogenesis. HDGF also plays a role in cell-cell interaction and cell migration. HDGF is a growth factor that is involved in stimulating vascular smooth muscle cells (SMCs)proliferation during development and in disease. HDGF contains
a true bipartite nuclear localization sequence necessary for nuclear targeting. HDGF is sciential factor in stimulating DNA replication and cell proliferation of vascular smooth muscle cell.In this study,we used E. coli strain BL21 (DE3) to express the recombinant protein hepatoma derived growth factor(HDGF). To find out the optimal production conditions,we studied on the different temperature and fermentor to calculate all cost .

第一章 前言..............................................1
一、肝癌的形成........................................1
二、肝癌衍生生長因子(HDGF)............................2
三、基因重組大腸桿菌之醱酵培養與誘導特性..............5
第二章 材料和實驗方法....................................9
一、基因選殖..........................................9
二、發酵槽使用步驟....................................13
三、蛋白質純化步驟....................................15
四、蛋白質濃度分析....................................17
五、SDS-PAGE 電泳分析.................................18
第三章 結果 ............................................20
一、以1L錐型瓶及5L發酵槽所得之HDGF單位成本分析…..20
二、以IPTG及Nacl在不同溫度下induction所得之HDGF濃
度比較...........................................25
第四章 討論.............................................26
第五章 參考文獻.........................................31

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