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ÇóÖú¸÷λ´óÏÀ£¬ÀÏʦÈÃд¸öÑо¿¼Æ»®£¬¿ÉÊÇÎÞ´¦ÏÂÊÖ°¡£¬Ö®Ç°²»ÊÇѧ·Ö×ÓÉúÎïѧµÄ£¬Çë¸÷λ¸øÒ»µã½¨Ò飬Ñо¿¼Æ»®µÄÌá¸ÙÈçÏ£º Regarding to your application, can you write a research proposal relevant to Wnt pathway? I would propose the following topic for your research proposal: Developing cell-based assay for Wnt signaling. The outline will be as followed: 1. Construct a vector encoding Frizzled receptor (a GPCR). Please include the detailed information of choosing the vetor, designing the primers for PCR and the whole cloning process, etc. 2. Construct a vector encoding report gene containing TCF-LEF responsive element (luciferase report gene or GFP gene) Please include the detailed information of choosing the vetor, designing the primers for PCR and the whole cloning process, etc. 3. Generate a cell line stably expressing the above two genes. Please include the process of stable cell line generation. 4. Expressing Wnt protein in bacteria expression system with affinity tag (His tag or GST tag) and purification of Wnt proteins. Please include the detailed information of choosing the vetor, designing the primers for PCR, the whole cloning process and protein purification, etc. 5. Verify the cell-based assay and carry out the protein activation assay During the writing process, you could refer to the webpage http://www.stanford.edu/group/nusselab/cgi-bin/wnt/ and check out the relevant literature. Íò·Ö¸Ð¼¤£¡£¡ |
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