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| Gene Synthesis | The open reading frames (ORF) for the PPRV (China/tibet/geg/07-30) M and H genes were obtained from NCBI (GenBank Access No. FJ905304.1). Codons in both ORFs were optimized based on Spodoptera frugiperda (Sf) cells using an OptimumGene™ software to eliminate the rare codons. The codon-optimized H ORF was flanked by restriction sites SmaI/NheI (Fig. 1A). The codon-optimized M ORF was flanked by restriction sites NsiI/KpnI, and a SV40 polyA signal and a polh promoter were also included (Fig. 1B). Both resulting fragments were synthesized and cloned in pUC57-Simple plasmids by Genscript Inc., China. | Get A Quote |
Peste des petits ruminants virus (PPRV), an etiological agent of peste des petits ruminants (PPR), is classified into the genus Morbillivirus in the family Paramyxovirida. In this study, two full-length open reading frames (ORF) corresponding to the PPRV matrix (M) and haemagglutinin (H) genes underwent a codon-optimization based on insect cells, respectively. Two codon-optimized ORFs along with one native nucleocapsid (N) ORF were used to construct recombinant baculoviruses co-expressing the PPRV M, H and N proteins in insect cells. Analysis of Western blot, immunofluorescence, confocal microscopy and flow cytometry demonstrated co-expression of the three proteins but at different levels in insect cells, an... More
