Peptide-based polyclonal antibody against mosquito 14-3-3ζ recognizes 14-3-3 homolog from dipteran and lepidopteran insects

Autor: Yongseok Lee, Iksoo Kim, Yong Hun Jo, Sook Jae Seo, Sun Am Kim, Yeon Soo Han, Seung Han Oh, Heung Cheol Kim, In Seok Bang, Mi Young Noh
Rok vydání: 2009
Předmět:
Zdroj: Entomological Research. 39:129-134
ISSN: 1748-5967
1738-2297
DOI: 10.1111/j.1748-5967.2009.00214.x
Popis: The 14-3-3 proteins are known to play an important regulatory role in apoptosis, and various cell signaling cascades. However, no investigation on mosquito 14-3-3 has been reported. To investigate the role of 14-3-3 proteins in mosquito midgut cells undergoing apoptosis, we decided to take advantage of Anopheles gambiae genome data, and were able to find Ag14-3-3ζ cDNA and protein sequences from Ensembl (http://www.ensembl.org). Further in silico analysis using BLAST search revealed that Ag14-3-3ζ protein is a polypeptide of 248 amino acids, and shares high identity with 14-3-3ζ homologues from Aedes aegypti (100%), Drosophila melanogaster (96%) and Bombyx mori (93%). Due to the perfect match and high homology, we hypothesized that Ag14-3-3ζ peptide antibody may recognize 14-3-3ζ homologs from other anopheline mosquitoes and insects. We thus generated 14-3-3ζ polyclonal antibody against a unique region located in the C-terminal end of Ag14-3-3ζ after in silico epitope analysis. As expected, zoo-western blot analysis of 14-3-3 proteins revealed that a polyclonal antibody against Ag14-3-3ζ peptide recognizes 14-3-3 homologs from dipteran and lepidopteran insects. To our knowledge, this is the first report on polyclonal antibody production against mosquito 14-3-3ζ. The mosquito-based 14-3-3ζ antibody will be very useful for studying the functional characterization of 14-3-3ζ in the context of host–pathogen interactions in midgut and other immune cells.
Databáze: OpenAIRE
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