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. 2022 Feb 28;37(3):341–347. doi: 10.1016/j.virs.2022.02.009

Fig. 1.

Fig. 1

Development of the ura3-52-defective strain VL6-48B. A A diagram of the ura3-52 locus in VL6-48 and primer pairs for amplifying the bsd gene (URA3-BSD-S/A), genotyping (BSD-S/URA3-Ter), and sequencing (URA3-Pro) of the transgene. The PCR amplicon of bsd contains 58- and 57-bp homologous sequences surrounding the ura3-52 open reading frame (ORF) at its 5′- and 3′-termini. The primers for genotyping targeted the start codon and 184 bp downstream of the ura3-52 ORF. B The genomic content of transformed yeast was extracted and served as the template for genotyping PCR. The amplicon was then subjected to gel electrophoresis, and a 583-bp band was seen by ethidium bromide staining. C The genomic content of transformed yeast was extracted and subjected to sequencing using the primer URA3-Pro. The sequenced region consists of bsd at the center region and the ura3 surrounding sequence at the 5′- and 3′-ends.