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Optical sensing of plankton communities and dynamics
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Study of subcellular localization of dinoflagellate rhodopsin by transformation and expression in mammalian cells P-B2-01-S Minglei Ma* , State Key Laboratory of Marine Environmental Science(Xiamen University) Xinguo Shi, State Key Laboratory of Marine Environmental Science(Xiamen University) Kaidian Zhang, State Key Laboratory of Marine Environmental Science(Xiamen University) Senjie Lin, State Key Laboratory of Marine Environmental Science(Xiamen University) Presenter Email: 18850583965@163.com |
Rhodopsins are now found in all domains of life. Based on amino acid sequences, rhodopsin proteins are classified into two large groups: type I, found in animal and type II found in microbes including Bacteria, Archaea, and Eukarya. The latter type of rhodopsin has been reported to convert light into ATP directly through light-driven proton pump in the ocean. Rhodopsin genes similar to that encoding proton-pumping rhodopsin in bacteria have been found in dinoflagellates. However, the Subcellular localization and the function of dinoflagellate rhodopsin are still poorly understood. In order to address the gap of knowledge, we constructed expression plasmid for Prorocentrum donghaisense and Alexandrium fundyence rhodopsin genes to transfect 293HEK cells. Total RNA was extracted from the stable transformed cells and RT-PCR performed. Results indicate that both of the P. donghaiense and A. fundyense rhodopsin genes have been incorporated into 293T successfully. The results of Western Blot show that the rhodopsin has been expressed in 293T cells successfully. The results of Subcellular localization shows that the rhodopsin is targeted to membrane. We will focus on studying functions of the rhodopsin gene which is to examine intracellular pH under illumination. |
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