Table 1.

Primers used for construction and analysis of mutant BACs

Primer namePrimer sequence
Sce-GmR Fora5′-GACATGGATCCTAGGGATAACAGGGTAATAATTGACATAAGCCTGTTCGGTTCG-3′
Sce-GmR Reva5′-GTAGCTCTAGAGGCCGCGGCGTTGTGAC
L64I Gm Fwdb5′-AACAGGAGCTGTGTTTACACGAGCGCCAGCGCTATCGGGaaTaTTCGCCGCCCTCGCCCAGACGCTCGGATCCTAGGGATAACAGGG-3′
L64I Gm Revb5′-TGGCGATCTCCTCGGAGGGCGTCTGGGCGAGGGCGGCGAAtAttCCCCGATAGCGCTGGCGCTCTCTAGAGGCCGCGGCGTTG-3′
Gm mid For5′-GGTCGTGAGTTCGGAGACGTAGC-3′
Gm mid Rev5′-CACTACGCGGCTGCTCAAACC-3′
L64I unique For 5′-AACAGGAGCTGTGTTTACACGAGCG-3′
L64I unique Rev 5′-TGGCGATCTCCTCGGAGGGC-3′
UL31 test Fwd5′-TGCCCCTGGTGAAGACCAC-3′
UL31 test Rev5′-GCTACGGCGGAGGAAACTCG-3′
  • a Underlined sequences have homology to the Gmr cassette.

  • b Underlined sequences have homology to the UL31 gene. Lowercase indicates nucleotides altered to create the L64I mutation and an SspI restriction enzyme cleavage site. Sequences not underlined have homology to the SceI-Gmr cassette.