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In particular, we describe separate lines with human α7 and human α4β2 nicotinic acetylcholine receptors, mouse 5-HT3A sereceptorseceptors and a chimera of human α7/mouse 5-HT3A receptors.
We optimized the methodology in mammalian cells by using an engineered Tyr-RS, and a chimera of human and Bacillus stearothermophilus tRNATyr to heterologously express low-abundance GPCRs.
We adapted the amber suppression technology to achieve heterologous expression of low-abundance GPCRs in mammalian cell culture by combining an engineered tyrosine synthetase originally developed in a yeast system with a novel chimera of human and Bacillus stearothermophilus tRNATyr.
Recombinant human PEPD was generated in bacteria as described recently and was incubated at 0.04, 0.2 or 1 μM with 0.04 μM of ErbB2/ECD-Fc (a recombinant chimera of human ErbB2 ECD (Thr23-Thr652) and the Fc fragment of human IgG1) or 0.04 μM of Fc as a control.
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Using a set of chimeras of human and mouse SCARB2, we identified that the region containing residues 77 113 of human SCARB2 contributes significantly to JL2 binding.
Based on the sequence analysis, we constructed a serious of chimeras of human and mouse SCARB2 (Fig. 3C) to map the binding site of JL2.
This observation in patients has been substantiated in human/SCID mice chimeras of human prostate cancer tumorigenesis [ 73].
The improvement in stability of human NAT1 was based on comparing the substrate specificity and stability of a series of chimeras of human NAT1 and human NAT2 (Goodfellow et al., 2000).
Using chimeras of the human β2-adrenergic and human A2A adenosine receptors, we present the methodology and data for the initial selection of an expanded set of fusion partners for crystallizing GPCRs.
But alongside growing interest in the use of chimeras for human health there has been vocal political opposition in the West.
Nakauchi and colleagues have evaluated the chimera-forming ability of human pluripotent stem cells in vitro (Masaki et al. 2015).
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