Exploration of IRES Elements within the ORF of the Coxsackievirus B3 Genome

Objective This study aimed to identify internal ribosome entry sites(IRESs)in the open reading frame(ORF)of the Coxsackievirus B3(CVB3)genome.Methods The sequences of P1,P2,or P3 of the CVB3 genome or the truncated sequences from each antithymocyte globulin(ATG)to the end of the P1,P2,or P3 gene were inserted into the pEGFP-N1vector.After transfection,possible IRES-dependent green fluorescent protein(GFP)-fused proteins were detected by anti-GFP western blotting.The sequences of possible IRESs were inserted into specific Fluc/Rluc bicistronic vectors,in which the potential IRESs were determined according to the Fluc/Rluc activity ratio.Expression of Fluc and Rluc mRNA of the bicistronic vector was detected by RT-qPCR.Results After transfection of full length or truncated sequences of the P1,P2,or P3 plasmids,six GFPfused protein bands in P1,six bands in P2 and nine bands in P3 were detected through western blotting.Two IRESs in VP2(1461–1646 nt)and VP1(2784–2983 nt)of P1;one IRES in 2C(4119–4564 nt)of P2;and two IRESs in 3C(5634–5834 nt)and 3D(6870–7087 nt)of P3 were identified according to Fluc/Rluc activity ratio.The cryptic promoter was also excluded by RT-qPCR.Conclusion Five IRESs are present in the CVB3 coding region.

Construction of MicroRNA-Target Interaction Networks Based on MicroRNA Expression Profiles of HRV16-infected H1-HeLa Cells

In the present study we investigated the changes in miRNA levels inhuman rhinovirus 16(HRV16)-infected cells.A small RNA deep sequencing experiment was performed through next-generation sequencing.In total,53 differentially expressed miRNAs were confirmed by RT-q PCR,including 37 known mi RNAs and 16 novel miRNAs.Interaction networks between differentially expressed miRNAs and their targets were established by mir DIP and Navigator.The prediction results showed that QKI.