Characterization and Localization of Fluorescent Pseudomonas Cold Shock Protein(s) by Monospecific Polyclonal Antibodies

Abstract

Cold shock protein (CSP) from Pseudomonas fluorescens MTCC 103 and cold resistant protein (CRP) from its mutant CRPF8 of 14 and 35 kd, respectively were purified to homogeneity by HPLC. Polyclonal antibodies were raised against these proteins and the expression level was checked at different temperatures, i.e., 4, 10, 20, 30 and 37 C. Furthermore, morphological changes in P. fluorescens MTCC 103 and its mutant (CRPF8) were analyzed by transmission electron microscopy (TEM). Localization of CSP and CRP documented with immunoelectron microscopy, using colloidal gold particles conjugated with secondary antibodies being the probe were used. Nevertheless, the results of cytosolic localization of CSP and CRP were evident. Furthermore, the expression of CSP and CRP increased with decrease in temperature and the cell wall thickness of the mutant exhibited 2‐fold increase, thus facilitating low temperature survival.

Leveraging Pseudomonas Stress Response Mechanisms for Industrial Applications

“…The expression level of a cold shock protein from P. fluorescens MTCC 103 (MW 14 kd) and a cold resistant protein (MW 35kd) from the P. fluorescens mutant CRPF8 were studied over a range of temperatures from 37 • C down to 4 • C (Khan et al, 2003). Expression of the cold shock protein and the cold resistant protein increased in response to decreased temperature with the rate of induction of protein synthesis reaching its maximum at 10 • C. Eighteen Pseudomonas strains isolated from Antarctica were analyzed for the presence of cold shock proteins and cold accumulation proteins (Panicker et al, 2010).…”

Molecular insights into the ecology of a psychrotolerant Pseudomonas syringae

“…CspA, a well‐known cold shock protein, is implicated in the modulation of RNA secondary structure, thereby facilitating the process of transcription and translation under stress conditions (Keto‐Timonen et al ., 2016; Zhang et al ., 2018a). Pseudomonas species such as P. fluorescens , P. fragi , and P. koreensis , as well as P. syringae Lz4W, are known to produce cold shock proteins (Ray et al ., 1994c; Michel et al ., 1997; Khan et al ., 2003; Awasthi et al ., 2019). In P. syringae Lz4W, CspA is upregulated upon shifting down to 4°C, suggesting involvement of cold‐shock protein in maintenance of RNA structure at low temperature (Ray et al ., 1994c; Bisht et al ., 2014; Zhang et al ., 2018a; Awasthi et al ., 2019).…”