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Distribution of an Ankyrin‐repeat Protein on the Endoplasmic Reticulum in Arabidopsis
Author(s) -
Wei Liqin,
Li Yan
Publication year - 2009
Publication title -
journal of integrative plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.734
H-Index - 83
eISSN - 1744-7909
pISSN - 1672-9072
DOI - 10.1111/j.1744-7909.2008.00791.x
Subject(s) - endoplasmic reticulum , polyclonal antibodies , ankyrin repeat , arabidopsis , ankyrin , fusion protein , microbiology and biotechnology , biology , stim1 , biochemistry , antibody , gene , recombinant dna , genetics , mutant
There are many ankyrin‐repeat proteins in plant cells. However, the distribution and function of these proteins are mostly unclear. By reverse transcription‐polymerase chain reaction, a gene encoding an ankyrin‐like protein was cloned from Arabidopsis and named AtANK1 (GenBank accession no. NM_120340). The 6‐His‐tagged AtAnk1‐N fusion protein was affinity‐purified and its rabbit polyclonal antibody was obtained. Immuno‐blotting with the purified anti‐AtAnk1‐N polyclonal antibody revealed that the relative molecular weight of the AtANK1 protein was about 76 kDa. By immunofluorescence labeling and immuno‐gold labeling with the purified anti‐AtAnk1‐N polyclonal antibody, coupled with confocal and transmission electron microscopy observation, AtANK1 was found to be distributed on the membrane of the endoplasmic reticulum in Arabidopsis cells. Based on these results, we suggested that AtANK1 might be involved in endoplasmic reticulum‐related protein localization and sorting in plant cells.