Integration of E. coli aroG-pheA tandem genes into Corynebacterium glutamicum tyrA locus and its effect on L-phenylalanine biosynthesis

Figure 1 Pathway and primary regulations of phenylalanine biosynthesis in C. glutamicum←---indicates feedback inhibition, ---→indicates activation.

Figure 2 Construction of recombinant plasmids. E: EcoRI; P: PstI; H: Hind III; GA: P_BF-aroG-pheA-T fragment; GAK: P_BF-aroG-pheA-T-Km fragment.

Figure 3 Insertion of P_BF-aroG-pheA-T-Km fragment of pTGAK into chromosome tyrA locus of C. glutamicum by double-ex-change homologous recombination and detection of ligated DNA fragment Km-tyrA tail on chromosome by PCR ampli-fication with P3 and P5.

Figure 4 DNA detection of transformants by PCR amplifica-tion and PstI digested PCR products. Lane 1: control for PCR; lane 2: PstI digesting product of lane 3; lane 3: PCR detection of C. glutamicum KM; lane 4, PstI digesting product of lane 5; lane 5: PCR detection of C. glutamicum GAK; lane 6: Markers, EcoRI/Hind III digesting λ phage DNA.

Figure 5 Auxanogram detection of C. glutamicum (Tyr^-). MM: C. glutamicum GAK on MM; Tyr: C.glutamicum GAK on MM + Tyr; Phe: C.glutamicum GAK on MM + Phe; Trp: C.glutamicum GAK on MM + Trp.