FIG. 6.

Characterization of VLPs isolated from yeast cells containing the reporter plasmid pGPol(−)hisAI and the pJEFWTneo plasmid or the pJEFMutINneo plasmid. (A) VLP-associated RT activity. (B) Immunoblotting with anti-RT, anti-IN, and anti-Gag antibodies. The volumes of VLPs that were found to incorporate the same amount of dGTP into high-molecular-mass poly(dG) by the oligo(dG)/poly(rC) primer template assay were loaded onto the gel and probed with the antibodies. (C) Production of double-stranded cDNA in VLPs. The VLP DNA was digested with EcoRI or Eco91I (BstEII), separated on a 1% agarose gel, transferred to a hybond N⁺ membrane, and probed with ³²P-labeled DNA. The EcoRI-digested DNA was probed with a HIS-labeled fragment. The 2.3-kbp band is the Ty1-HIS digested DNA. The Eco91I-digested DNA was probed with a neo-labeled fragment which hybridizes with a 5.0-kbp Ty1-neo digested cDNA and cross-hybridizes with the 5.2-kbp Ty1-HIS digested cDNA.