Proteomic Analysis of Lactococcus garvieae Cultivated under Planktonic and Biofilm Conditions and Identification of Biofilm Formation-Related Immunogenic Proteins
Date Issued
2015
Date
2015
Author(s)
Wang, Ting-Chieh
Abstract
Lactococcus garvieae is an aquatic pathogen that causes disease in a wide range of aquatic species all over the world. It is generally believed that latent bacterial infections and resulting antibiotic resistance are caused by the in vivo formation of biofilms. Moreover, previous studies have revealed that biofilm formation causes a change in protein expression and the antigen profile available for host-recognition. In view of this, proteomic and immunoproteomic analysis were used to compare the proteomes of L. garvieae grown either as a biofilm or in the planktonic state. Proteomic analysis using two-dimensional gel electrophoresis (2-DE) revealed varying amounts of protein in nine protein spots, among which seven were up-regulated and two were down-regulated in the biofilm form when compared to the planktonic controls. Among the up-regulated proteins, four of them (glmU, gtaB, rmlB, and rfbC) participated in extracellular polymeric substances (EPS) synthesis, which is the main component of biofilm. In the immunoproteomic analysis, polyclonal rabbit antibodies raised against biofilm and planktonic bacteria reduced biofilm formation by L. garvieae, while the anti-biofilm serum showed stronger inhibition of biofilm formation. The sera were later used to perform 2-DE Western blot assays on whole cell protein extracted from both biofilm and planktonic forms. Two immunoreactive proteins specifically recognized by anti-biofilm sera were selected, among which a moonlighting protein, arginine deiminase (AD), was considered to be associated with biofilm formation, and thus may be a potential vaccine candidate against biofilm infections.
Subjects
Lactococcus garvieae
biofilm
two-dimensional gel electrophoresis
immunoproteomic
arginine deiminase
SDGs
Type
thesis
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