Results 101 to 110 of about 1,116 (265)

Mycobacterial cell division arrest and smooth‐to‐rough envelope transition using CRISPRi‐mediated genetic repression systems

open access: yesFEBS Open Bio, EarlyView.
CRISPRI‐mediated gene silencing and phenotypic exploration in nontuberculous mycobacteria. In this Research Protocol, we describe approaches to control, monitor, and quantitatively assess CRISPRI‐mediated gene silencing in M. smegmatis and M. abscessus model organisms.
Vanessa Point   +7 more
wiley   +1 more source

C2α‐carbanion‐protonating glutamate discloses tradeoffs between substrate accommodation and reaction rate in actinobacterial 2‐hydroxyacyl‐CoA lyase

open access: yesFEBS Open Bio, EarlyView.
Enzymes of the 2‐hydroxyacyl‐CoA lyase group catalyze the condensation of formyl‐CoA with aldehydes or ketones. Thus, by structural adaptation of active sites, practically any pharmaceutically and industrially important 2‐hydroxyacid could be biotechnologically synthesized. Combining crystal structure analysis, active site mutations and kinetic assays,
Michael Zahn   +4 more
wiley   +1 more source

Single‐molecule DNA flow‐stretch assays for high‐throughput DNA–protein interaction studies

open access: yesFEBS Open Bio, EarlyView.
We describe an optimised single‐molecule DNA flow‐stretch assay that visualises DNA–protein interactions in real time. Linear DNA fragments are tethered to a surface and stretched by buffer flow for fluorescence imaging. Using λ and φX174 DNA, this protocol enhances reproducibility and accessibility, providing a versatile approach for studying diverse ...
Ayush Kumar Ganguli   +8 more
wiley   +1 more source

Analysing the significance of small conformational changes and low occupancy states in serial crystallographic data

open access: yesFEBS Open Bio, EarlyView.
This protocol paper outlines methods to establish the success of a time‐resolved serial crystallographic experiment, by means of statistical analysis of timepoint data in reciprocal space and models in real space. We show how to amplify the signal from excited states to visualise structural changes in successful experiments.
Jake Hill   +4 more
wiley   +1 more source

Screening and epitope characterization of Nidogen‐2‐specific nanobodies

open access: yesFEBS Open Bio, EarlyView.
Camel immunization and phage display were employed to generate high‐affinity VHH nanobodies against Nidogen‐2. After library construction, biopanning, ELISA screening, sequencing, and recombinant expression, selected nanobodies were purified and characterized, leading to the preliminary exploration of a nanobody‐based sandwich ELISA for specific ...
Jianchuan Wen   +9 more
wiley   +1 more source

Introducing Intel(R) SAT Solver.

open access: yes, 2022
We introduce Intel(R) SAT Solver (IntelSAT) - a new open-source CDCL SAT solver, written from scratch. IntelSAT is optimized for applications which generate many mostly satisfiable incremental SAT queries. We apply the following Incremental Lazy Backtracking (ILB) principle: in-between incremental queries, backtrack only when necessary and to the ...
openaire   +3 more sources

Proteasomal degradation of intracellularly expressed Amblyomin‐X limits suicide gene therapy potential in melanoma cells

open access: yesFEBS Open Bio, EarlyView.
This study explores the feasibility of expressing the antitumoral protein Amblyomin‐X through a suicide gene therapy approach and investigates its intracellular fate after gene delivery. Although the gene is efficiently expressed, melanoma cells rapidly degrade the Amblyomin‐X protein via proteasome activity.
Victor Dal Posolo Cinel   +4 more
wiley   +1 more source

Decreased cold‐sensing function of the transient receptor potential channel TRPM8 from tailed amphibians

open access: yesFEBS Open Bio, EarlyView.
Despite frogs avoiding low temperatures, examination of four salamander species revealed that none avoided cold and all possessed cold tolerance. Functional analysis of TRPM8, a cold sensor, showed that all salamander TRPM8s had lost their cold sensitivity.
Tadahiro Sawao   +3 more
wiley   +1 more source

A Verified SAT Solver Framework with Learn, Forget, Restart, and Incrementality. [PDF]

open access: yesJ Autom Reason, 2018
Blanchette JC   +3 more
europepmc   +1 more source

Rapid screening of staphylokinase protein variants using an unpurified cell‐free expression system

open access: yesFEBS Open Bio, EarlyView.
An unpurified cell‐free protein synthesis (CFPS) platform enables rapid functional screening of staphylokinase variants. Direct plasminogen‐activation assays performed in microplate format provide real‐time activity readouts, allowing rapid identification and ranking of variants with improved or reduced fibrinolytic activity without protein ...
Maria Tomková   +3 more
wiley   +1 more source

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