Journal ArticleDOI
Characterization and expression of the Hex 110 gene encoding a glutamine-rich hexamerin in the honey bee, Apis mellifera.
Márcia Maria Gentile Bitondi,Adriana Mendes do Nascimento,Adriana D. Cunha,Karina R. Guidugli,Francis M. F. Nunes,Zilá Luz Paulino Simões +5 more
TLDR
Semiquantitative and quantitative RT-PCR with specific primers allowed for an analysis of mRNA levels during worker bee development and under different physiological conditions, which revealed a glutamine-rich polypeptide with a predicted molecular mass of 112.2 kDa and pI of 6.43.Abstract:
An N-terminal amino acid sequence of a previously reported honey bee hexamerin, HEX 110 [Danty et al., Insect Biochem Mol Biol 28:387-397 (1998)], was used as reference to identify the predicted genomic sequence in a public GenBank database. In silico analysis revealed an ORF of 3,033 nucleotides that encompasses eight exons. The conceptual translation product is a glutamine-rich polypeptide with a predicted molecular mass of 112.2 kDa and pI of 6.43, which contains the conserved M and C hemocyanin domains. Semiquantitative and quantitative RT-PCR with specific primers allowed for an analysis of mRNA levels during worker bee development and under different physiological conditions. Concomitantly, the abundance of the respective polypeptide in the hemolymph was examined by SDS-PAGE. Hex 110 transcripts were found in high levels during the larval stages, then decreased gradually during the pupal stage, and increased again in adults. HEX 110 subunits were highly abundant in larval hemolymph, decreased at the spinning-stage, and remained at low levels in pupae and adults. In 5th instar larvae, neither starvation nor supplementation of larval food with royal jelly changed the Hex 110 transcript levels or the amounts of HEX 110 subunit in hemolymph. In adult workers, high levels of Hex 110 mRNA, but not of the respective subunit, were related to ovary activation, and also to the consumption of a pollen-rich diet.read more
Citations
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Journal ArticleDOI
Validation of reference genes for gene expression studies in the honey bee, Apis mellifera, by quantitative real-time RT-PCR
TL;DR: Three software-based approaches were used and four candidate genes were selected as suitable genes for normalization in A. mellifera to evaluate the suitability of these genes as endogenous controls to standardize qRT-PCR results.
Journal ArticleDOI
CpG methylation in the hexamerin 110 gene in the European honeybee, Apis mellifera
TL;DR: Observations suggest that the methylation of the Hex110 gene is regulated at the developmental stage and in a caste-dependent manner.
Journal ArticleDOI
The four hexamerin genes in the honey bee: structure, molecular evolution and function deduced from expression patterns in queens, workers and drones.
Juliana Martins,Francis M. F. Nunes,Alexandre S. Cristino,Zilá Luz Paulino Simões,Márcia Maria Gentile Bitondi +4 more
TL;DR: In this paper, the authors explored the hexamerin genes found in the honey bee genome by exploring their structural characteristics, expression profiles, evolution, and functions in the life cycle of workers, drones and queens.
Journal ArticleDOI
Insulin receptor substrate influences female caste development in honeybees
TL;DR: Proteomic differences between irs gene knockdowns and controls are revealed, including levels of hexamerin 110, a storage protein connected to natural caste differences, which is a key adaptor protein in IIS.
Journal ArticleDOI
Standard methods for physiology and biochemistry research in Apis mellifera
Klaus Hartfelder,Márcia Maria Gentile Bitondi,Colin S. Brent,Karina Rosa Guidugli-Lazzarini,Zilá Luz Paulino Simões,Anton Stabentheiner,Erica D. Tanaka,Ying Wang +7 more
TL;DR: Sensitive techniques for the quantification of juvenile hormone levels circulating in haemolymph, as well as its synthesis by the corpora aliata are described and a standard method for the detection and analysis of certain biogenic amines of interest is presented.
References
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