Peptide-based polyclonal antibody against mosquito 14-3-3ζ recognizes 14-3-3 homolog from dipteran and lepidopteran insects |
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Authors: | Sun Am KIM Mi Young NOH Yong Hun JO Seung Han OH Iksoo KIM Yong Seok LEE Heung Cheol KIM Sook Jae SEO In Seok BANG Yeon Soo HAN |
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Affiliation: | Department of Agricultural Biology, College of Agriculture and Life Science, Chonnam National University, Gwangju, Korea; Department of Parasitology, College of Medicine and Frontier Inje Research for Science and Technology, Inje University, Busan, Korea; US Army, 5th Medical Detachment, 18th Medical Command, Seoul, APO AP 96205-0020, Korea; Division of Applied Life Science, Gyeongsang National University, Jinju, Korea; Department of Biological Science and the Research Institute for Basic Sciences, Hoseo University, Asan, Korea |
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Abstract: | 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. |
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Keywords: | antibody cross-reactivity insect 14-3-3ζ Korea mosquito |
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