expression and localization of an hsp70 protein in the microsporidian encephalitozoon cuniculihsp70蛋白的表达和定位在microsporidian encephalitozoon cuniculi.pdfVIP

Hindawi Publishing Corporation International Journal of Microbiology Volume 2010, Article ID 523654, 6 pages doi:10.1155/2010/523654 Research Article Expression and Localization of an Hsp70 Protein in the Microsporidian Encephalitozoon cuniculi Carrie E. Jolly, Cory A. Leonard, and J. Russell Hayman Department of Microbiology, James H. Quillen College of Medicine, East Tennessee State University, Box 70579, Johnson City, TN 37614, USA Correspondence should be addressed to J. Russell Hayman, hayman@ Received 19 April 2010; Accepted 22 June 2010 Academic Editor: Robert P. Gunsalus Copyright © 2010 Carrie E. Jolly et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Microsporidia spore surface proteins are an important, under investigated aspect of spore/host cell attachment and infection. For comparison analysis of surface proteins, we required an antibody control specific for an intracellular protein. An endoplasmic reticulum-associated heat shock protein 70 family member (Hsp70; ECU02 0100; “C1”) was chosen for further analysis. DNA encoding the C1 hsp70 was amplified, cloned and used to heterologously express the C1 Hsp70 protein, and specific antiserum was generated. Two-dimensional Western blotting analysis showed that the purified antibodies were monospecific. Immunoelectron microscopy of developing and mature E. cuniculi spores revealed that the protein localized to internal structures and not to the spore surface. In spore adherence inhibition assays, the anti-C1 antibodies did not inhibit spore adherence to host cell surfaces, whereas antibodies to a known surface adhesin (EnP1) did so.