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. 2021 Oct 4;13(10):1991. doi: 10.3390/v13101991

Figure 1.

Figure 1

Assembly of a full-length cDNA clone of PDCoV CHN-HN-2014. (A) Schematic diagram of the intermediate plasmid pBAC-M-PDCoV. A gene fragment obtained by multi-step fusion PCR, containing CMV promoter, a partial N-terminal region of PDCoV genome (nt 1–375), a partial C-terminal region of PDCoV genome (nt 23,365–25,421), a 27-nt poly(A) tail, the hepatitis deltavirus (HDV) sequence, and the bovine growth hormone (BGH) termination sequences, was cloned into the backbone vector pBeloBAC11 between the restriction sites ApaLI and HindIII. The critical restriction sites and their relative positions in the genome of the PDCoV strain CHN-HN-2014 are indicated. (B) Five contiguous genomic overlapping overhangs were amplified and assembled into pBAC-M-PDCoV by homologous recombination. (C) Schematic diagram of the full-length cDNA clone of CHN-HN-2014 in the pBeloBAC11 vector, referred to as pBAC-CHN-HN-2014.