Seven different mutants were constructed harboring, separately or in combination, three different genetic modifications to Synechocystis' metabolic network.
To examine if the lower Vmax is due to a decreased expression, chimeras were constructed harboring green fluorescent protein (GFP) fused to the C-terminus of the Hxt36 and the Hxt36-N367I mutant and expressed in the DS68625 strain.
In addition, we generated a GST-fusion construct harboring the otherwise unstable UCS domain (UNC-45GST-UCS).
A similar observation was made for the construct harboring roughly half the hydrophobic sequence of the gp41 TM segment (Fig. 5a, compare lanes 6 and 8).
Each shRNA construct harbors a specific barcode of 18 nucleotides (nt) adjacent to the shRNA sequence that facilitates subsequent identification of shRNAs.
When expressed in an A. fumigatus wild type strain, the construct harboring only the sixth HAMP domain exerts a strong dominant negative effect on the native TcsC.
These repressions were completely abrogated when miR-208b and/or miR-499 were co-transfected with a luciferase reporter construct harboring a mutated miR-208b/miR-499 binding site in the Col25a1 3′UTR.
Briefly, HCT116 cells were seeded at 2 × 104 cells/well on 96-well plates and transfected with 100 ng of luciferase construct harboring NF-κB response elements using Lipofectamine 3000 (Invitrogen, Thermo Fisher Scientific).
In these experimental conditions, we analyzed TPA action upon endogenous ALAS mRNA levels, as well as the promoter activity of a fusion reporter construct, harboring the TPA-responsive region of ALAS gene driving chloramphenicol acetyl transferase gene expression.
In a cell-free assay, GloSensor showed increased luminescence in proportion to the amount of FCV protease, while no signal change was observed when the construct harboring the mutant cleavage motif was used.
For rice inplanta transformation pre-germinated seed embryonic axis was wounded with a fine needle and the PsAKR1 construct harboring agrobacterium was infected.
