Fluorescent co-localization of PTS1 and PTS2 and its application in analysis of the gene function and the peroxisomal dynamic in Magnaporthe oryzae
文献类型： 外文期刊
作者： Wang, Jiao-yu 2 ; Wu, Xiao-yan 3 ; Zhang, Zhen 2 ; Du, Xin-fa 2 ; Chai, Rong-yao 2 ; Liu, Xiao-hong 1 ; Mao, Xue-qin 2 ;
作者机构： 1.Zhejiang Univ, Coll Agr & Biotechnol, State Key Lab Rice Biol, Hangzhou 310029, Zhejiang, Peoples R China
2.Zhejiang Acad Agr Sci, Hangzhou 310021, Zhejiang, Peoples R China
3.NW Agr & Forest Univ, Coll Plant Protect, Yangling 712100, Peoples R China
关键词： green fluorescence;biological metabolism;biological roles;fluorescent fusions;peroxisomal targeting signals;pre-penetration processes
期刊名称：JOURNAL OF ZHEJIANG UNIVERSITY-SCIENCE B （ 影响因子：3.066； 五年影响因子：3.057 ）
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收录情况： SCI
摘要： The peroxisomal matrix proteins involved in many important biological metabolism pathways in eukaryotic cells are encoded by nucleal genes, synthesized in the cytoplasm and then transported into the organelles. Targeting and import of these proteins depend on their two peroxisomal targeting signals (PTS1 and PTS2) in sequence as we have known so far. The vectors of the fluorescent fusions with PTS, i.e., green fluorescence protein (GFP)-PTS1, GFP-PTS2 and red fluorescence protein (RFP)-PTS1, were constructed and introduced into Magnaporthe oryzae Guy11 cells. Transformants containing these fusions emitted fluorescence in a punctate pattern, and the locations of the red and green fluorescence overlapped exactly in RFP-PTS1 and GFP-PTS2 co-transformed strains. These data indicated that both PTS1 and PTS2 fusions were imported into peroxisomes. A probable higher efficiency of PTS1 machinery was revealed by comparing the fluorescence backgrounds in GFP-PTS1 and GFP-PTS2 transformants. By introducing both RFP-PTS1 and GFP-PTS2 into Delta mgpex6 mutants, the involvement of MGPEX6 gene in both PTS1 and PTS2 pathways was proved. In addition, using these transformants, the inducement of peroxisomes and the dynamic of peroxisomal number during the pre-penetration processes were investigated as well. In summary, by the localization and co-localization of PTS1 and PTS2, we provided a useful tool to evaluate the biological roles of the peroxisomes and the related genes.
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