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By contrast, robust ATP-dependent Cl− currents were recorded in P274R iPSC-RPE complemented with WT hBest1-GFP expressing from a BacMam baculoviral vector (Fig. 5a, b), validating this system for studying ATP-dependency of hBest1.
We can solve this dilemma by inserting the cloned genes into the chromosome rather than expressing from a plasmid.
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This contrasts to previous studies in which the constitutive hacA was expressed from a highly-expressed promoter [ 43] or expressed from the pyrG locus [ 30].
These were cloned then expressed from a His tag expression plasmid and, following purification on nickel columns, identities were confirmed by Western blot analysis.
RacB could also be inducibly expressed, whereas the protein could not be expressed from a constitutive promoter, presumably because expression at high levels is lethal.
In two prior studies, human AS3MT was expressed from a cDNA clone and purified after heterologous expression in E. coli.
The dual-expression system is comprised of codon-optimized Cullin3 and Rbx1 genes co-expressed from a single pET-Duet-1 plasmid.
MAVS, a regulator of antiviral innate immunity, is expressed from a bicistronic mRNA encoding a second protein, miniMAVS.
It is a view expressed from a position of relative strength.
Similarly, the CRISPR-resistant phage D1811 (Fig. 1c) titres increased when its Acr was expressed from a plasmid.
The tetracycline repressor and tRNA suppressor (Glu) are expressed from a single G418 selectable vector, while a gene engineered to contain a stop codon is expressed from a separate hygromycin selectable vector.
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