Fig. 1.

Expression of the endogenous TAR1 gene can be detected at the transcript level, but not the protein level. a Detection of the endogenous TAR1 transcript by reverse transcriptase PCR (RT-PCR). All reactions were carried out in duplicate using two different RNA preparations. Lanes 1 and 2 show the results of RT-PCR on samples treated with DNase and with no RT enzyme included in the cDNA synthesis step (control for contaminating genomic DNA); lanes 3 and 4 show PCR products generated from genomic DNA in samples treated with RNase (control for size of expected RT-PCR product); and lanes 5 and 6 show a product amplified from the endogenous TAR1 transcript by RT-PCR in the complete absence of genomic DNA. b Western blot using a Tar1p peptide antibody. Lanes 1 and 2 are duplicate protein samples from a wild type strain transformed with empty vector; lanes 3 and 4 are duplicate protein samples from a strain transformed with a vector expressing Tar1p from the constitutive TEF1 promoter