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Characterization and Localization of Fluorescent Pseudomonas Cold Shock Protein(s) by Monospecific Polyclonal Antibodies
Author(s) -
Khan Mahejibin,
Bajpai Virendra Kumar,
Anasari Shaheen Akhtar,
Kumar Anil,
Goel Reeta
Publication year - 2003
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2003.tb03456.x
Subject(s) - polyclonal antibodies , immunoelectron microscopy , pseudomonas fluorescens , biology , mutant , microbiology and biotechnology , antibody , protein a , primary and secondary antibodies , fluorescence microscope , fluorescence , biochemistry , bacteria , immunology , gene , physics , genetics , quantum mechanics
Abstract Cold shock protein (CSP) from Pseudomonas fluorescens MTCC 103 and cold resistant protein (CRP) from its mutant CRPF 8 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 (CRPF 8 ) 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.