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. 2018 Sep 25;9:2250. doi: 10.3389/fmicb.2018.02250

FIGURE 1.

FIGURE 1

Plasmids construction. (A) RNA-launched infectious clones absent/present poly(A) tail or 3′ UTR were established. Besides, the mutated RNA-launched infectious clones possessed various length of poly(A) tail were also constructed and named pR-DHAV-An (n = 0, 5, 10, 15, 20, 25, 30, and 40). (B) Construction of the DHAV-1 monocistronic reporter system. The plasmid pDHAV-3′UTR-A25 contains the following elements 5′ to 3′ in a pcDNA3.1/V5-His A vector: the CMV immediate early promoter, a T7 promoter, the entire DHAV-1 5′ UTR (nucleotides 1–626, strain LY0801), and the FLuc gene and the entire DHAV-1 3′ UTR followed by a poly(A) tail containing 25 adenines. pDHAV-Δ3′UTR-ΔA25, or pDHAV-3′UTR-ΔA25, or pDHAV-Δ3′UTR-A25 were obtained from pDHAV-3′UTR-A25 to remove 3′ UTR plus poly(A) tail, or poly(A) tail, or 3′ UTR, respectively. The mutated monocistronic reporter plasmids possessed various length of poly(A) tail was constructed and named as pDHAV-3′UTR-An (n = 0, 5, 10, 15, 20, 25, 30, and 40).