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Pro247-Ser variant was constructed based on homology modelling and rational design.
An additional D8S1179 reverse primer specific for the variant was constructed resulting in the recovery of the null allele.
For this purpose, a Gag variant was constructed encoding a 105 kDa protein fusion containing Gag at its N-terminus (to ensure in vivo VLP assembly and GFP encapsulation), followed by an 11 amino acid T7 epitope tag (for immunological detection) and a factor Xa cleavage site to release mature GFP from Gag (Figure 6A).
The H107AH108A variant was constructed as previously reported.
To maximize the destabilizing interactions, the R141E/Y288A/A306F variant was constructed.
For this reason, the R103K variant was constructed to investigate the possibility that a lost charge-based stabilization was responsible for the increased KM in the R103Q variant.
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For the quantitative real-time PCR assay, two standard curves specific for each splice variant were constructed.
A set of nine catalytically diverse P450 BM3 variants was constructed and tested toward the native substrate-inspired fluorogenic substrate 12- 4-trifluoromethylcoumarin-7-yloxy dodecanoic 12- 4-trifluoromethylcoumarin-7-yloxy dodecanoic 12- 4-trifluoromethylcoumarin-7-yloxy dodecanoic 12- 4-trifluoromethylcoumarin-7-yloxy dodecanoic 12- 4-trifluoromethylcoumarin-7-yloxy dodecanoic
A library of enzyme variants was constructed by staggered extension process (StEp) using the genes that code for the l-ASNases from Erwinia chrysanthemy (ErL-ASNase) and Erwinia carotovora (Ecal-ASNase) and screened using activity assays.
Subsequently, an alternative splicing graph of potential splice variants was constructed.
A network of MHC variants was constructed with the Neighbor-Net method [ 75] by using Splits-tree4 ver. 4.11.3 [ 76].
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