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Inverse PCR for Site-Directed Mutagenesis
Inverse PCR is a powerful tool for the rapid introduction of desired mutations at desired positions in a circular double-stranded DNA sequence. In...
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Design and Production of a Novel Anti-PD-1 Nanobody by CDR Grafting and Site-Directed Mutagenesis Approach
Programmed cell death protein-1 (PD-1) is a membrane protein expressed on the surface of activated T-cells, B-cells, natural killer cells, dendritic...
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Site-Directed Mutagenesis Method Mediated by Cas9
This study presents an in vitro CRISPR/Cas9-mediated mutagenic (ICM) system that allows rapid construction of designed mutants or site-saturation...
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A new halotolerant xylanase from Aspergillus clavatus expressed in Escherichia coli with catalytic efficiency improved by site-directed mutagenesis
Daily agro-industrial waste, primarily cellulose, lignin, and hemicellulose, poses a significant environmental challenge. Harnessing...
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A simplified Gibson assembly method for site directed mutagenesis by re-use of standard, and entirely complementary, mutagenesis primers
BackgroundSite-directed mutagenesis (SDM) is a key method in molecular biology; allowing to modify DNA sequences at single base pair resolution....
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Study of the Mechanism of Coenzyme Specificity of Phenylacetone Monooxygenase from Thermobifida fusca by Site-Directed Mutagenesis
AbstractPhenylacetone monooxygenase from Thermobifida fusca (EC 1.14.13.92, PAMO) belongs to the Baeyer–Villiger family of monooxygenases and...
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Site-Directed Mutagenesis Protocol to Determine the Role of Amino Acid Residues in Polycomb Group (PcG) Protein Function
Site-directed mutagenesis (SDM) is a technique in molecular biology and protein engineering that is widely used to determine the significance of...
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Improved catalytic activity of a novel aspartate kinase by site-directed saturation mutagenesis
This study aimed to improve the catalytic activity of aspartate kinase (AK), the first key rate-limiting enzyme in the aspartic acid metabolism...
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Molecular Modeling and Optimization of Type II E.coli l-Asparginase Activity by in silico Design and in vitro Site-directed Mutagenesis
IntroductionL-asparaginase (also known as L-ASNase) is a crucial therapeutic enzyme that is widely used in treatment of ALL (acute lymphoblastic...
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Cleavable Cys labeling directed Lys site-selective stapling and single-site modification
Site-selective modification of peptide/protein is a vital approach to disclose post-translational modifications (PTMs) and plays a crucial role in...
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Site-directed mutagenesis of the quorum-sensing transcriptional regulator SinR affects the biosynthesis of menaquinone in Bacillus subtilis
BackgroundMenaquinone (MK-7) is a highly valuable vitamin K 2 produced by Bacillus subtilis . Common static metabolic engineering approaches for...
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Facile Assembly of Combinatorial Mutagenesis Libraries Using Nicking Mutagenesis
Combinatorial mutagenesis is a method where multiple user-defined mutations are encoded at defined positions in a sequence. Combinatorial mutagenic...
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A High-Specific-Activity L-aspartate-α-Decarboxylase from Bacillus aryabhattai Gel-09 and Site-Directed Mutation to Improve Its Substrate Tolerance
L-aspartate-α-decarboxylase (ADC) can recognize L-aspartic acid specifically and catalyze the decarboxylation of L-aspartic acid to β-alanine. In...
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Improvement of thermostability and catalytic efficiency of glucoamylase from Talaromyces leycettanus JCM12802 via site-directed mutagenesis to enhance industrial saccharification applications
BackgroundGlucoamylase is an important industrial enzyme in the saccharification of starch into glucose. However, its poor thermostability and low...
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Identification of Aggregation-Prone and Gatekeeper Residues in Bacterial Amyloids Using Site-Directed Mutagenesis and Flow Cytometry
Bacterial functional amyloids are remarkable examples of how amyloid aggregation can be kept under control and even leveraged to perform diverse...
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Site-directed mutagenesis in the P-domain of calreticulin transacylase identifies Lys-207 as the active site residue
In silico-docking studies from previous work have suggested that Lys-206 and lys-207 of calreticulin (CR) play a pivotal key role in its...
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Improvement in the catalytic performance of a phenylpyruvate reductase from Lactobacillus plantarum by site-directed and saturation mutagenesis based on the computer-aided design
To enhance the specific activity and catalytic efficiency ( k cat / K m ) of an NADH-dependent Lp PPR, its directed modification was performed based on the...
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Structural Flexibility of Tau in Its Interaction with Microtubules as Viewed by Site-Directed Spin Labeling EPR Spectroscopy
Tau is a microtubule-associated protein that belongs to the Intrinsically Disordered Proteins (IDPs) family. IDPs or Intrinsically Disordered Regions...
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GeneORator: An Efficient Method for the Systematic Mutagenesis of Entire Genes
Directed evolution is a powerful tool for the rapid improvement of a target protein toward a desired fitness criteria, such as activity, specificity,...
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Domain Shuffling and Site-Saturation Mutagenesis for the Enhanced Inhibitory Potential of Amaranthaceae α-Amylase Inhibitors
Amaranthaceae α-amylase inhibitors (AAIs) are knottin-type proteins with selective inhibitory potential against coleopteran α-amylases. Their small...
