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. 2003 Jul;77(14):7830–7842. doi: 10.1128/JVI.77.14.7830-7842.2003

FIG. 1.

FIG. 1.

Schematic presentation of the construction of hybrid FFV-FCV vectors. (A) Parental GFP expression vector pCF-Bet-Gfp (29). FFV genes and LTRs (subdivided into the U5, R, and U3 regions) are represented by open boxes. The inserted GFP sequence is shown as a shaded box inserted into the deletion in the U3 region of the 3′ LTR. Below is shown amplification of the FCV E sequences E14, E23, and E24 with different combinations of primers 1 to 4. Primers 1 and 2 contain a terminal NheI site, and primers 3 and 4 contain a NotI site. (B) Hybrid FFV-FCV clones (pCF-FCVx) carrying a deletion in the U3 sequence of the 3′ LTR. The FCV E inserts (E14, E23, and E24) are represented by shaded boxes. The restriction sites and primers (horizontal arrows) used to regenerate the U3 are shown. (C) Chimeric FFV-FCV clones (pCF-FCVx-U3) with the reconstructed U3. The different FCV E inserts (E14, E23, and E24) are represented by shaded boxes. Below, PCR primers (horizontal arrows) and a DNA probe used to detect and characterize reisolated vector genomes are presented. (D) Different FCV E inserts are shown in the single-letter code. The numbers above the FCV E sequence refer to the FCV-KS20 capsid protein gene sequence (11).