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Example sentences for: prs

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Each strain was transformed with a plasmid harboring the respective wild type gene in pRS316 (pSW1268 or pSW1269; see below) [ 66 ] and sporulated to isolate haploid secΔ deletion strains with the p SEC/URA3/CEN plasmid.
URA3 was amplified as a ~ 1.1 kb fragment from pRS306 [ 32 ] . Before TAR cloning, the vector was linearized with Bam HI to expose the B1 targeting sequence and the URA3 sequence placed at the end of the transgene-specific hook.
The PCR product was digested with SacI and XbaI and cloned into the SacI/XbaI sites of both pRS315 (pSW1264) and pRS316 (pSW1268) [ 66 ] . Plasmids with wild type SEC23 were constructed in the same manner using specific oligonucleotides (5' oligo sec23F5 with 5'-ATGAATATCTAGACCAGGGTGCC-3'; 3' oligo sec23-3 with 5'CCTTGGATCCGTAGTAAAGGCCACGCAG-3') and digestion of the PCR product with XbaI and BamHI to yield p SEC23/LEU2 (pSW1266) and p SEC23/URA3 (pSW1269).
The 1.8 kb SacI-SalI fragment of the original library clone, containing only the 5' half of NrG1 without the zinc fingers, was cloned into the SacI-SalI sites of pRS426 to create pHZ56.
Dengue-2 virus cDNA cloned in the yeast shuttle vector pRS424, linearized by excision of a short Bam H1 fragment was transfected into competent [ 45 ] S. cerevisiae YPH857 (kindly provided by Barry Falgout, (CBER/FDA), along with the appropriate PCR fragment spanning the desired deletion.