Selective Pressure Incorporation
Selective Pressure Incorporation - Related Videos
11:56
Antimicrobial Peptides Produced by Selective Pressure Incorporation of Non-canonical Amino Acids
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Cited by 15 •
2018
The protocol presents the Escherichia coli-based selective pressure incorporation of non-canonical amino acids (ncAAs) into the lactococcal antimicrobial peptide nisin. Its properties can be changed during recombinant expression via substitution with desired ncAAs in defined growth media. Resulting changes in bioactivity are mapped by growth inhibition assays and fluorescence microscopy.
11:51
Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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Cited by 7 •
2018
Synthetic biology enables the engineering of proteins with unprecedented properties using the co-translational insertion of non-canonical amino acids. Here, we presented how a spectrally red-shifted variant of a GFP-type fluorophore with novel fluorescence spectroscopic properties, termed "gold" fluorescent protein (GdFP), is produced in E. coli via selective pressure incorporation (SPI).
07:26
Synthesis and Microdiffraction at Extreme Pressures and Temperatures
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Cited by 5 •
2013
The laser heated diamond anvil cell combined with synchrotron micro-diffraction techniques allows researchers to explore the nature and properties of new phases of matter at extreme pressure and temperature (PT) conditions. Heterogeneous samples can be characterized in situ under high pressure by 2D mapping and combined powder, single-crystal and multigrain diffraction approaches.
08:54
Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method
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Cited by 175 •
2009
The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. This tail-cuff system uses Volume Pressure Recording (VPR) to measure the blood pressure by determining the tail blood volume.
08:53
Natural Selection - Student Protocol
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2019
Simulating Natural Selection ExpandTo begin, cut the white, black, and gray pipe cleaners into quarters, which should produce a total of 100 pieces that are 3 inches long. Then, cut the green pipe cleaners in the quantities and lengths specified in the video, taking care to minimize waste. This approximates a normal distribution of lengths and should result in an additional 100 pieces total. NOTE: One 12-inch pipe cleaner should yield multiple lengths, for example, a 5-inch piece and a 7-inch...
FAQs
The auxotrophic host creates the essential control point: withholding the required natural amino acid removes its ordinary supply, while adding a structural analog provides an alternative substrate. If the host’s existing aminoacyl-tRNA synthetase recognizes the analog, it can attach it to the corresponding tRNA, allowing translation to incorporate the substituted residue at selected codons.
Recognition by the existing aminoacyl-tRNA synthetase determines whether the analog can enter translation through the host’s normal tRNA-handling machinery. The analog must resemble the withheld natural amino acid sufficiently for the relevant synthetase to recognize and attach it to the corresponding tRNA. This recognition step connects the supplied compound to protein synthesis.
Replacing a natural amino acid with a noncanonical analog can introduce chemical behavior unavailable from the standard amino-acid set. The resulting proteins may show altered reactivity or stability, or gain labeling potential. These changes make the approach useful when bioengineers want to examine how a protein’s chemical composition influences its structure or function.
A typical setup begins with an amino-acid-auxotrophic host, followed by withholding the required natural amino acid and supplying a structural analog. The host’s existing aminoacyl-tRNA synthetase must recognize that analog and attach it to the corresponding tRNA. Translation can then place the analog at selected codons, producing a protein for downstream study or engineering.
Researchers can use the resulting proteins to investigate structure and function, especially when altered reactivity, stability, or labeling potential is informative. The same outputs can support biosensor development and the engineering of therapeutic or industrial biomolecules. Thus, the method connects controlled amino-acid substitution with both mechanistic protein studies and useful engineered products.
In bioengineering, the method expands the chemical functions available to cells and engineered biomolecules beyond those provided by natural amino acids alone. That expanded chemical range gives researchers a way to pursue protein designs with changed reactivity, stability, or labeling potential, supporting applications such as biosensors and therapeutic or industrial biomolecule engineering.