[Objective]The aim of the study is to establish the microRNA library of Rhipicephalus microplus,to identify the species of microR-NA including the library,to analyze the secondary and tertiary structure of Vg-2 protein regulated by miR-275,and to predict the signal pep-tides,transmembrane domain,phosphorylation and glycosylation sites,and to dominant epitopes of Vg-2 protein.[Method]The adult female R.microplus was used as sample.The microRNA library of R.microplus was established by using the high-throughput sequencing,and the software miRDP and miRBase database were used to identify the species of microRNA including this library.The online software EXPASY,PRABI and SWISS-MODEL were used to analyze the physical and chemical properties and to infer the secondary and tertiary structure of Vg-2 protein regulated by miR-275.The online software SignalP 5.0,TMHMM,NetPhos 3.1 and NETCGlyc 1.0 were used to detect the signal peptides,transmembrane domain,phosphorylation and glycosylation sites of Vg-2 protein,and the software ABCpred Prediction,Scratch,IEDB and NetCTL were used to predict the dominant antigen epitopes of Vg-2 protein in B and T cells.[Result]Results of our study showed that a microRNA library including total of 383 microRNAs was established,12 of which microRNAs were identified and annotated,371 of which microRNAs were newfound,and 85 microRNAs had the rich contents among these microRNAs.The length of the Vg-2 gene se-quence of R.microplus was 5040 bp,and this gene totally encoded 1679 amino acids.Vg-2 protein was a hydrophilic acid protein that was 51.98 kDa in molecular size,and its theoretical isoelectric point(pI)was 6.08.Vg-2 protein used βsheet as its major structural compo-nents.There were 268 phosphate sites and 20 glycosylation sites,but was no signal peptide and transmembrane domain in the amino acids sequence of Vg-2 protein.Vg-2 protein had 58 B cell dominant antigen epitopes and 8 T cell dominant antigen epitopes.[Conclusion]There are 383 microRNAs in the established microRNA library of the adult bloody female R.microplus,and the Vg-2 protein regulated by miR-275 is a hydrophilic acid protein with dominant antigen epitopes.Vg-2 protein with the good antigenicity and immunogenicity is an ideal target for the future research and development of vaccine against R.microplus.
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