Amplification of TMUV-prM Gene

The prM gene specific primers were designed based on the published ORF sequence of TMUV-TC2B. The specific primers (F, 5′-CTGAAGCTTGGAAACTATAATGGCAG-3′;R, 5′-GCTGTACGCTGGGGCAAT T-3′) were designed according to the published ORF sequence of the TMUV prM gene (GenBank:{"type":"entrez-nucleotide","attrs":{"text":"MH764605","term_id":"1811395401","term_text":"MH764605"}}MH764605), and the prM gene was amplified using the cDNA of the TMUV-TC2B strain as a template. The PCR reaction conditions included predenaturation at 95°C for 3 min, denaturation at 95 °C for 30 s, annealing at 55 °C for 30 s, extension at 72 °C for 30 s, 32 cycles, and finally extension at 72°C for 5 min. The PCR products were observed by electrophoresis in a 1% agarose gel.