Search

Objective: To sequence the complete genome of CVS-11 strain and establish a reverse genetic system of CVS-11 to further study its pathogenic mechanism, virulence genes and antigenic sites., Methods: We amplified12 fragments covering the complete genome of the CVS-11 strain by RT-PCR, and then cloned to pEASY-Blunt vector for sequencing the complete genome of CVS-11. We analyzed single restriction enzyme sites of the full length cDNA of the CVS-11 strain by DNAMAN and designed 4 pairs of specific primers. We amplified the full-length cDNA of CVS-11 by RT-PCR. We obtained four fragments and cloned into pcDNA3. 1. We named the full-length cDNA plasmid pcDNA3. 1-CVS-11. We also cloned helper plasmids pcDNA3.1-N, P, L and G expressing N, P, L and G protein of CVS-11 strain, respectively. We co-transfected NA cells with the full-length plasmid and four helper plasmids. We identified the supernatant of the transfected and then passaged NA cells by immunofluorescence staining and RT-PCR and found the recombinant virus rCVS-11 rescued successfully., Results: Sequencing results showed that the complete genome of CVS-11 was composed of 11 927 nucleotides. The complete genome of CVS-11 encoded 5 structure proteins and gene array was the same as other reported rabies viruses. We successfully constructed a reverse genetic system of CVS-11, namely the full length plasmid pcDNA3. 1-CVS-11 and 4 help plasmids pcDNA3. 1-N, P, L, G and rescued the rCVS-11 from a full-length infectious cDNA clone., Conclusion: The reverse genetic system of the CVS-11 strain laid the foundation for future studies on rabies virus.

Aptamers are a group of artificial oligonucleotides identified by exponential enrichment system evolution technology (Selective expansion of ligands by exponential enrichment, SELEX). Aptamers have been widely used in basic research, clinical diagnostics, and nano-technology. In this article we will introduce the technology of aptamer and summarize its applications in medical microbiology.