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. 2016 Nov 10;6:35543. doi: 10.1038/srep35543

Figure 4. Analysis of the Thph1 and Thph2 gene expression levels in the mutant strains.

Figure 4

(A) PCR confirmation of the ΔThph1 knock-out mutant using four pairs of primers: Group 1, amplification of the hph gene. Group 2, amplification of the Thph1 gene. Group 3, amplification of right border flanking fragments of T-DNA. Group 4, amplification of the 5′ flanking sequence of Thph1 (WT, wild-type strain; P, plasmid; ΔThph1, knock-out mutant of ΔThph1; M, DNA ladder). (B) PCR confirmation of the ΔThph2 knock-out mutant using four pairs of primers: Group 1, amplification of the hph gene. Group 2, amplification of the Thph2 gene. Group 3, amplification of the right border flanking fragments of T-DNA. Group 4, amplification of the 5′ flanking sequence of Thph2 (WT, wild-type strain; P, Plasmid, ΔThph2, knock-out mutant of ΔThph2; M, DNA ladder). (C) PCR verification of the ΔThph1&Thph2 double knock-out mutant. Lane 1, amplification of the hph gene using the WT DNA as the template. Lane 2, amplification of the G418 gene using the WT DNA as the template. Lane 3, amplification of the hph gene using the mutant DNA as the template. Lane 4, amplification of the G418 gene using the mutant DNA as the template. Lane 5, amplification of the Thph1 gene using the WT DNA as the template. Lane 6, amplification of the Thph2 gene using the WT DNA as the template. Lane 7, amplification of the Thph1 gene using the mutant DNA as the template. Lane 8, amplification of the Thph2 gene using the mutant DNA as the template. Lane 9, amplification of the flanking segment of the Thph1 gene using the WT DNA as the template. Lane 10, amplification of the flanking segment of the Thph2 gene using the WT DNA as the template. Lane 11, amplification of the flanking segment of the Thph1 gene using the mutant DNA as the template. Lane 12, amplification of the flanking segment of the Thph2 gene using the mutant DNA as the template.