Identification and Partial Characterization of an L-Tyrosine Aminotransferase (TAT) from Arabidopsis thaliana
Figure 6
(a) Functional complementation assay. Functional complementation was tested using the E. coli mutant DL39. The plasmids used were pBAD33 and pBAD33 + At5g36160. The bacteria were grown to an OD of 1.0 measured at 600 nm, the strains were serially diluted to 10-1, 10-2, 10-3, and 10-4 using 0.85% (w/v) saline, and 5 L was replica plated on medium with or without L-Tyrosine and L-Phenylalanine supplemented with 0.2% (w/v) arabinose as described in Section 2. It should be noted that 50 individual transformants were replica plated and tested using the complementation assay. All 50 bacteria transformed with the empty vector were unable to grow on media lacking L-Tyrosine or L-Phenylalanine while all 50 transformants expressing the plant enzyme were able to grow on media lacking L-Tyrosine and L-Phenylalanine. For presentation purposes, only one control and one experimental transformant were used in the diagram. (b) shows the growth curve of the DL39 strain transformed with either pBAD33 (triangle) or pBAD33 + At5g36160 (circle) in M9 medium lacking L-Tyrosine and L-Phenylalanine supplemented with 0.2% (w/v) arabinose. (c) shows the summary of the functional complementation assay of the plant enzyme ability to complement the E. coli triple mutant on M9 medium.
