Construction of Full-Length Genomes-Modified cDNA Clone for Rabies Virus Strain SRV_9

In order to obtain full-length genomes-modified cDNA clone for rabies virus strain SRV9 and to deeply study genomic characteriitics of SRV9 and construct the new virus vector and several pairs of primers were designed according to the full-length genomic sequence of SRV9 published on GenBank by using the total RNA extracted from rabies virus as a template.Five DNA fragments covering the entire genome 11 928 bp were amplified,and inserted into vector and were determined for sequeneing.After identification of the sequence,Ψ region of genome was designed with a lack of primers before the human cytochrome c gene was cloned into such a region,and cytoplasmic region of glycoprotein gene was also deleted by primers designed ahead.Hammerhead ribozyme sequence was inserted into the head region of full-length genomes and hepatitis delta virus ribozyme sequence was added into the end region of full-length genomes with the help of primers.Fragments were all cloned into pcDNA3.1(+) vector after sequenced correctly.These five fragments were ligated by using amplified fragments that contain their own restriction enzyme sites to obtain the full-length genomes-modified plasmid clone pcDNA3.1-SRV9,which would contribute to the foundation of infectious clone for rabies virus.