A rhodopsin-like protein in Cyanophora paradoxa: gene sequence and protein immunolocalizatioN

Authors: A.M. Frassanito, L. Barsanti, V. Passarelli, V. Evangelista, P. Gualtieri
Journal: Cellular and Molecular Life Sciences
DOI: 10.1007/s00018-009-0225-x
Abstract:
Here, we report the DNA sequence of the rhodopsin gene in the alga Cyanophora paradoxa (Glaucophyta). The primers were designed according to the conserved regions of prokaryotic and eukaryotic rhodopsin-like proteins deposited in the GenBank. The sequence consists of 1,272 bp comprised of 5 introns. The correspondent protein, named Cyanophopsin, showed high identity to rhodopsin-like proteins of Archea, Bacteria, Fungi, and Algae. At the N-terminal, the protein is characterized by a region with no transmembrane alpha-helices (80 aa), followed by a region with 7alpha-helices (219 aa) and a shorter 35-aa C-terminal region. The DNA sequence of the N-terminal region was expressed in E. coli and the recombinant purified peptide was used as antigen in hens to obtain polyclonal antibodies. Indirect immunofluorescence in C. paradoxa cells showed a marked labeling of the muroplast (aka cyanelle) membrane.

Keywords: Rhodopsin-like protein, Cyanophora paradoxa, Muroplast, Cyanelle, Antibody labeling

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