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Figure 1 | BMC Biotechnology

Figure 1

From: Incorporation of a lambda phage recombination system and EGFP detection to simplify mutagenesis of Herpes simplex virus bacterial artificial chromosomes

Figure 1

Schematic diagram of allele replacement strategy using vector pKO5. (A) Features of a typical allele replacement vector include a positive selection marker for antibiotic resistance marker (zeo), a temperature-sensitive origin of replication (ts-ori), a negative selection marker (sacB) and flanking sequences to targeting cross-over at hypothetical gene D. Incubation at non-permissive temperature (43°C) and selection on Zeocin/Chloramphenicol lead to co-integration through homologous recombination (single cross-over). (B) Diagram of the resultant single-crossover recombinant BAC and the two possible forms following resolution. Incubation at permissive temperature (30°C) and selection on Zeocin/Sucrose lead to resolution of the co-integrate, excision of the plasmid through homologous recombination (double cross-over).

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BMC Biotechnology

ISSN: 1472-6750

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