Bioinformatic and gene expression pattern of adenylat kinase gene in Nosema ceranae
The objective of this study is to analyze the physicochemical property and molecular characteristics of the adenylate kinase(ADK)in Nosema ceranae,and detect the expression feature of NcADK during the N.ceranae infection of Apis mellifera ligustica worker,enriching relevant information of and providing a basis for further functional investigation.The physicochemical properties,signal peptides,phosphorylation sites,and secondary and tertiary structures of NcADK were predicted and analyzed by using related bioinformatics software.MEME software and Batch CD-Search tool were employed to predict the structural domains and conserved motifs in ADK proteins in N.ceranae and 7 other species.The phylogenetic tree was constructed by Mega 11.0 software based on the amino acid sequences of ADK proteins.RT-qPCR was performed to determine relative expression level of NcADK in N.ceranae during the infection process.The results indicated that the NcADK CDS contained 540 nucleotides encoding 179 amino acids;the molecular weight of NcADK was approximately 20.66 kDa,the molecular formula was C903H1488N258O278S8,the lipophilicity coefficient was 100.61,and the isoelectric point was 6.83;NcADK included 29 negatively charged amino acids and 29 positively charged amino acids;NcADK could be meantime localized in the cytoplasm,mitochondria,nucleus,vesicles,and peroxisomes;NcADK contained 20 phosphorylation sites while lacked typical signal peptide;NcADK included 88 a-helices,24 extended strands,17 β-turns,and 50 random coils,and it shared 100％sequence homology with the template A0A0F9WEU7.1.A;the same one structural domains and the same 5 conserved motifs were indetified in NcADK and the ADK of Hamiltosporidium tvaerminnensis,Encephalitozoon intestinalis,Anncaliia algerae and Vittaforma corneae;NcADK and Nosema apis ADK were clustered into one clade on the evolutionary tree;as compared with that at 1 dpi(day post inoculation),the expression level of NcADK was up-regulated at 2 dpi without significant difference(P＞0.05),whereas the expression level of NcADK was significantly up-regulated at 3 dpi and 4 dpi(P＞0.05).These findings clarify the physicochemical property and molecular characteristics of NcADK,and suggested that NcADK was a potential hydrophilic and intracellular protein,ADK in different microsporidian species were highly conserved,the homology between NcADK and ADK in N.apis was the highest,and the expression of NcADK was activated at 3 dpi and 4 dpi.
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