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Table 3 Description of primers that were used in the present work for genome-based deletion of the catB gene (NCgl2318) and integration of the tuf-promoter for overexpression of the catA gene (NCgl2319) in Corynebacterium glutamicum

From: Metabolic engineering of Corynebacterium glutamicum for the production of cis, cis-muconic acid from lignin

Name Sequence Use AT [°C]
ΔcatB TS1 FW 5′-TTAACAATTGGGATCCTCTAGACCCACTCCAGAGCATTGGGGTGTTT-3′ Amplification of TS1; TS1 FW+ overlap to pClik, TS 1 RV+ overlap to TS2 FW 55
ΔcatB TS1 RV 5′-AGGTCCAATCGGCACCGCAGCAT-3′
ΔcatB TS2 FW 5′-GGATGCTGCGGTGCCGATTGGACCCTCAAGCGCCTTCATGGTCTCAAC-3′ Amplification of TS2; TS2 FW+ overlap to TS1 RV, TS 2 RV+ overlap to pClik 57
ΔcatB TS2 RV 5′-GCAGCCCGCTAGCGATTTAAATCCCTGGGCACCCATGGCGCCCAG-3′
Pef-tu-catA-TS1 FW 5′-AATTGGGATCCTCTAGACCCACGATTCCCTCTTCAGGGCG-3′ Amplification of TS1; TS1 FW+ overlap to pClik, TS1 RV+ overlap to Pef-tu 61
Pef-tu-catA-TS1 RV 5′-CATTCGCAGGGTAACGGCCAGTCATACTCCTAGAGACTTTGGGGG-3′
Pef-tu-catA-Peftu FW 5′-AAAGTCTCTAGGAGTATGACTGGCCGTTACCCTGCGAATG-3′ Amplification of Peftu; Peftu FW+ overlap to TS1 RV, Peftu RV+ overlap to TS2 FW 62
Pef-tu-catA-Peftu RV 5′-ATCTGTTCAGCTGAAGTCATTGTATGTCCTCCTGGACTTCGTGG-3′
Pef-tu-catA-TS2 FW 5′-GAAGTCCAGGAGGACATACAATGACTTCAGCTGAACAGATCG-3′ Amplification of TS2; TS2 FW+ overlap to Pef-tu, TS2 RV + overlap to pClik 57
Pef-tu-catA-TS2 RV 5′-CCGCTAGCGATTTAAATCCCTAGTATCCGTCCTCATCTGCG-3′
PR154 5′-ATTGTCTGTTGTGCCCAGTCATAG-3′ Validation of vector pClik int sacB (including respective insert) 53
PR318 5′-AATAATAGTGAACGGCAGGT-3′
  1. AT annealing temperature