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Cloning of ferritin gene AmFer3HCH and its response to temperature stress in the Italian honeybee,Apis mellifera ligustica (Hymenoptera:Apidae)
Pages: 1029-1039
Year: Issue:  9
Journal: Acta Entomologica Sinica

Keyword:  Apis mellifera ligusticaferritingene cloningtemperature stressexpression analysis;
Abstract: 【Aim】This study aims to clone and identify the ferritin gene Am Fer3 HCH from Apis mellifera ligustica,to analyze its tissue expression changes in response to temperature stress,and to explore the role of ferritin in encountering the extreme temperature stress.【Methods 】 The cDNA sequence of Am Fer3 HCH gene was amplified from A.m.ligustica by PCR,and its amino acid sequence was identified by bioinformatic tools.The expression levels of this gene in the thorax,midgut,and fat body of A.m.ligustica worker adults under temperature stress( 40℃ and 5℃) were detected by real-timequantitative PCR.【Results】A ferritin gene named Am Fer3 HCH was cloned from A.m.ligustica by PCR and its cDNA fragment is 507 bp in length.Its nucleotide sequence has 100% identity to that of the Fer3 HCH gene( GenBank accession no.: GB55416) in A.m.ligustica genome in GenBank database.Am Fer3 HCH was identified as a ferritin heavy-chain subunit and clustered in one phyletic clade with other heavy-chain ferritins.The amino acid sequence of Am Fer3 HCH has a typical ferroxidase activity center,ferrihydrite nucleation center and iron channel as in most ferritin family members.Real-time quantitative PCR analysis showed that the expression levels of Am Fer3 HCH in the thorax,midgut and fat body of A.m.ligustica worker adults after brief exposure to high temperature stress( 40℃ for 0-48 h)presented a tendency of upregulation first,then downregulation and upregulation again as compared to the control group.However,after the adults were exposed to brief low temperature stress( 5℃ for 0-6 h),the expression level of Am Fer3 HCH was up-regulated first,and then down-regulated in the midgut and fat body,but showed a falling tendency in the thorax.【Conclusion】Am Fer3 HCH is a heavy-chain ferritin and plays an important role in response to extreme temperature stress in A.m.ligustica.
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