Recombination between the two GFP copies by either intra- or inter-chromatid HR (Figure 1A), but not single strand annealing, can generate wild type GFP (wtGFP), which is readily quantified by flow cytometry.
They were further backcrossed 6 times to commercially available C57Bl (Jackson Laboratories, Bar Harbor, ME) and then intercrossed to generate wild type, heterozygous and homozygous plasminogen deficient mice.
In addition, mouse apoE appears to be more prone to lead to parenchymal plaque formation while human apoE4 is more prone to lead to CAA formation in mice that generate wild type human Aβ peptide [ 10, 24].
Finally, we generated wild-type or Tnfa /– gBT-I TRM cells in Tnfa /– mice before B16.gB challenge, as in Fig. 4c.
We generated wild-type and GC plants expressing the genetically-encoded GRX-roGFP2 probe, which specifically reports the ratio between the reduced and oxidized forms of glutathione (GSH/GSSG 38.
We also generated wild-type and mutant single-cell clones that innervated the DL1 glomerulus (Fig. S2E, F).
To express MsrB1 in mammalian cells, the following constructs were generated: wild-type and cysteine (Cys) mutant cDNA of mouse MsrB1 (NM_013759) corresponding to Form 1 3'-UTR were amplified from pET28a MsrB1-Sec and pET28a MsrB1-Cys as templates that were previously constructed in the laboratory.
To accomplish this, we first generated wild-type and PD mutant LRRK2 constructs with and without the WD40 domain (WD40 lacking constructs are referred to as "ΔWD"; Fig. 2A), and confirmed that all molecules exhibited similar stability and expressed at similar levels (Supplemental Figure S1).
The generated wild-type and mutated full-length molecular clones were used to produce stocks of wild-type and mutant viruses (termed HIV-1, HIV-1-AP-1 totmut, HIV-1-AP-1 totmutHIV-1-AP-1 totmutHIV-1-AP-1#3mut HIV-1-AP-1 totmut2mut).
PC generated wild-type and mutant yeast two-hybrid constructs.
