The constructs for RBDWT and mutants E484K, L452R/E484Q, and N440K were all designed as single genes containing each protein of interest fused to an N‐terminal µ‐phosphatase signal peptide and a C‐terminal hexahistidine tag. An Avitag™ BAP sequence followed by an EPEA C‐tag replaced the 6xHis tag in the ACE2‐expressing construct. Gene synthesis was performed by Twist Biosciences, which, using the NotI/XbaI restriction sites, also inserted the sequences into its standard vector pTwist‐CMV‐BetaGlobin‐WPRE‐Neo.
Ptwist
Manufactured by Twist Bioscience
Sourced in United States
The PTwist is a precision laboratory instrument designed for accurate and reliable DNA synthesis. It utilizes Twist Bioscience's proprietary technology to generate high-quality synthetic DNA sequences. The core function of the PTwist is to efficiently synthesize custom DNA molecules, providing researchers with a versatile tool for various applications in the life sciences.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ptwist
1
Recombinant SARS-CoV-2 Spike Protein Expression
2
Antibody Plasmid Construction and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The R175H antibody plasmids encoding the heavy-chain and light-chain15 (link) were synthesized, and the antibody was purified by Syd Labs, Inc. The single-chain variable fragment (scFv) was obtained from a previous study.17 (link) Assembly of antibody heavy- and light-chain DNAs into a mammalian expression vector pTwist (Twist Bioscience, South San Francisco, CA, USA) or gWIZ (Aldevron, Fargo, ND, USA) was performed to construct pR175H-mAb or pR175H/mCD3-BsAb. The plasmids were transformed into competent Escherichia coli cells and propagated in Luria–Bertani (LB) broth supplemented with 100 μg/mL ampicillin (Teknova, Inc., Hollister, CA, USA) or 50 μg/mL kanamycin (Teknova, Inc.). Plasmids were purified from DH5α cells grown overnight using an endotoxin-free ZymoPURE™ II Plasmid Maxiprep Kit (Zymo Research, Irvine, CA, USA).
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