FIG. 1.

Genomic organization of SqLCV-E and construction of expression vectors for wild-type and mutant AR1. (A) Restriction map of SqLCV-E. A HindIII site was introduced in the B component by site-directed mutagenesis. (B) Expression vectors for in vitro transcription and translation. To construct pGEM-AR1, the DdeI-DdeI fragment was excised from SqLCV-A_E, blunted, and ligated into SmaI-digested pGEM7Zf+. EagI and NheI sites on pBSKS-AR1 were introduced by site-directed mutagenesis for construction of AR1^Δ9-99 and AR1^Δ122-251, respectively. T7, T7 promoter. (C) Construction of GST fusions for expression in bacterial cells. The two EagI sites (italicized and underlined) were introduced by site-directed mutagenesis to construct pGST-AR1^Δ98-251 and pGST-AR1^Δ164-251, respectively. See text for details.