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Immuno-gold localization of hepatotoxins in cyanobacterial cells

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Abstract

A polyclonal antibody against the potent hepatotoxic cyclic peptide microcystins and nodularins was used in conjunction with immuno-gold labelling to localize the toxins in three strains of cyanobacteria. Ultrastructurally, there were no major differences between unicellular Microcystis aeruginosa strain PCC 7820 (toxin-producing strain) and M. aeruginosa strain UTEX 2063 (nontoxin-producing strain), except that M. aerginosa PCC 7820 cells has a sheath. The thickness of the sheath was about 12nm and was distinguishable from the cell wall at the ultrastructural level only when the specimen was stained en bloc with uranyl acetate. Microcystins and nodularin were found in M. aeruginosa PCC 7820 and Nodularia spumigena strain L-575 respectively, but not in nontoxic M. aeroginosa UTEX 2063. In M. aeruginosa PCC 7820 cells, microcystin was found primarily in the thylakoid area and nucleoid, with smaller amounts in the cell wall and sheath. Only nonspecific labelling was found in other cellular inclusions, such as polyhedral bodies, cyanophycin granules and membrane-limited inclusions. In strain N. spumigena L-575, nodularin was found in both vegetative cells and heterocysts with a distribution similar to that in M. aeruginosa PCC 7820.

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Abbreviations

Adda :

3-Amino-9-methoxy-10-phenyl-2,6,8-trimethyl-deca-4 (E), 6 (E)-dienoic acid

ELISA :

Enzyme-linked immunosorbant assay

PBS :

Phosphatebuffered saline

PBST :

PBS containing Triton X-100

PCC :

Pasteur Culture Collection

UTEX :

University of Texas Culture Collection

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Shi, L., Carmichael, W.W. & Miller, I. Immuno-gold localization of hepatotoxins in cyanobacterial cells. Arch. Microbiol. 163, 7–15 (1995). https://doi.org/10.1007/BF00262197

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