Establishment of Cell Lines with GFPTax Fusion Proteins Expressionand Its Effects on Cellular Growth

• Main Author: 孫承溥
• Other Authors: 董馨蓮
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/47183926674505823999

碩士 === 國立臺灣大學 === 微生物學研究所 === 90 === HTLV-I Tax was fused with green fluorescence protein (GFP) to examine its effect on cell growth and response to cytotoxic agents. Using transient transfection experiment, the fusion construct GFPTax was able to stimulate NF-κB-directed transcription as Tax was in G2-2 cell lines, microscopic observation revealed that GFPTax distributed mainly around nuclear membrane (both inside and outside) in punctuated forms. However, in Jurkat cell lines GFPTax accumulated mainly in the nucleolus regions. A retrovirus-based doxycycline inducible system was adopted to generate cell clones in Hep G2 with variable expression levels of GFPTax, with highest in G2-2-GFPTax7, medium in G2-2-GFPTax4 , and lowest (or none) in G2-2-GFPTax11 andG2-2-GFPTax6 . Colony-formation ability decreased in G2-2-GFPTax7. G2-2-GFPTax7 cells showed lower sensitivity toward anti-cancer agent, 5-Fu, as compared to G2-2-GFPTax11 cells. Flow cytometric analysis indicated that granularity of G2-2-GFPTax7 cells was higher. The cell cycle in G2-21-GFPTax7 cells was not different from that in the other cells with lower expression levels of GFPTax.