碩士 / 國立中山大學 / 海洋生物科技暨資源學系研究所 / 101 / The expressed sequence tags, (ESTs) derived from the developing tilapia brain, were established in our lab. There are 1092 transcripts. A transcript out of 1092 transcripts of ESTs was identified as a partial fragment of Transducer of ErbB2,1(Tob1). The unique gene, Tob1 encode a member of Btg/Tob protein family and play a role in cell growth. Bioinformatics were searched for Genbank: EE724041.1(Tob1), which is a gene with 1020 bp of open reading frame(ORF), and deduced as 339 amino acid of protein sequence. The pEGFPN1-Tob1 plasmid constructed by inserting tob1 into pEGFPN1 vector, was transfected into mouse neuroblastoma cell (neuron-2a cell, N2a). The distribution of Tob1 expressed in pEGFPN1-Tob1 transfected N2a cell was observed by confocal microscopy. This result shows that Tob1 protein is distributed in the cytoplasm of N2a. Tob1 protein was expressed by BL21(Escherichia coli), a prokaryotic system, and purified with Ni-NTA affinity chromatography. The effects of Tob1 protein on cell proliferation were investigated. These results show that the proliferation of neuro-2a cells is affected by Tob1.
| Identifer | oai:union.ndltd.org:TW/101NSYS5277014 |
| Date | January 2013 |
| Creators | Cheng-hsin Wang, 王正欣 |
| Contributors | Ching-Lin Tsai, 蔡錦玲 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 58 |
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