systematic analysis of fkbp inducible degradation domain tagging strategies for the human malaria parasite plasmodium falciparum系统分析fkbp诱导降解域标记策略人类恶性疟原虫.pdfVIP

Systematic Analysis of FKBP Inducible Degradation Domain Tagging Strategies for the Human Malaria Parasite Plasmodium falciparum 1,2 2,3 1 ˜ 1 Mauro Ferreira de Azevedo , Paul R. Gilson , Heloisa B. Gabriel , Roseli F. Simoes , 4,5 4,5 2,5 1 Fiona Angrisano , Jacob Baum , Brendan S. Crabb , Gerhard Wunderlich * ˆ ´ ˜ ˜ 1 Departamento de Parasitologia, Instituto de Ciencias Biomedicas, Universidade de Sao Paulo, Sao Paulo, Brasil, 2 The Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Victoria, Australia, 3 Monash University, Melbourne, Victoria, Australia, 4 The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia, 5 University of Melbourne, Victoria, Australia Abstract Targeted regulation of protein levels is an important tool to gain insights into the role of proteins essential to cell function and development. In recent years, a method based on mutated forms of the human FKBP12 has been established and used to great effect in various cell types to explore protein function. The mutated FKBP protein, referred to as destabilization domain (DD) tag when fused with a native protein at the N- or C-terminus targets the protein for proteosomal degradation. Regulated expression is achieved via addition of a compound, Shld-1, that stabilizes the protein and prevents degradation. A limited number of studies have used this system to provide powerful insight into protein function in the human malaria parasite Plasmodium falciparum. In order to bett