碩士 / 國立清華大學 / 生物資訊與結構生物研究所 / 93 / The original objectives of this research were to examine the distribution of lipid transfer protein (LTP) and monitor the transport pathway of LTP in plants by means of fusion to enhanced green fluorescence protein (EGFP). This research was done using the mungbean VrLtp1 cDNA fused to EGFP. Plasmids p35S��-VrspLTP1-EGFP-NOS, p35S��-EGFP-NOS, p35S��-NOS and p35S��-Vrsp1-EGFP-NOS were constructed and transiently expressed in protoplasts prepared from tobacco mesophyll and soybean suspension cells. The transient expression was detected by fluorescence microscopy and measured using a multilabel counter fluorometer. The green light emission by the protoplasts transformed with p35S��-EGFP-NOS was detected under a fluorescent microscope. Interestingly, protoplasts transformed with p35S��-VrspLTP1-EGFP-NOS exhibited bright yellowish green fluorescence with deformed shape. Using the Multilabel Counter Victor3 to measure EGFP expression, the EGFP counts in protoplasts transformed with p35S��-VrspLTP1-EGFP-NOS and p35S��-EGFP-NOS were similar, about 3.2 and 3.4 fold of that of the control. Our results indicate that the p35S��-VrspLTP1-EGFP-NOS fusion protein was expressed in protoplasts and the fusion protein can be used for further study on localization of lipid transfer protein.
| Identifer | oai:union.ndltd.org:TW/093NTHU5112015 |
| Date | January 2005 |
| Creators | 張楓瀅 |
| Contributors | 林彩雲 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 61 |
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