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This study investigated five mutations introduced into the TEV protease enzyme to improve its solubility. Following the mutations, a technique involving centrifugation was used to evaluate the solubility of each variant. The variants were concentrated ,samples were collected at specific time points, and then the protein concentrations were measured to assess their solubility. | This study investigated five mutations introduced into the TEV protease enzyme to improve its solubility. Following the mutations, a technique involving centrifugation was used to evaluate the solubility of each variant. The variants were concentrated ,samples were collected at specific time points, and then the protein concentrations were measured to assess their solubility. | ||
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Revision as of 17:34, 10 September 2024
C-TEV
Part Description
This is known as the tobacco etch virus which is very selective for cleaving proteins at particular amino acid sequences and has been modified to have better qualities like greater heat stability, decreased autolysis (self-cleavage) and also allows researchers to precisely cleave the tag off, leaving behind the receptor in its unmodified form. In our model, TEV was divided into N-terminal and C-terminal fragments. So the C-terminal fragment was grafted onto dCAS9(C) to generate TEV(C)-dCas9(C).
Usage
This is known as the tobacco etch virus which is very selective for cleaving proteins at particular amino acid sequences and has been modified to have better qualities like greater heat stability, decreased self-cleavage and also allows researchers to precisely cleave the tag off, leaving behind the receptor in its unmodified form. In our model TEV was divided into N-terminal and C-terminal fragments. So the C-terminal fragment was grafted onto dCAS9(C) to generate TEV(C)-dCas9(C).
A)They observed ccp1 in proteins of brain, spinal cord and peripheral nerves.They confirmed that by detection of extra band in HEK293 transfected with ccp1-YFP fusion proteins and absence of signals in pcd mouse brain B)They found relation of ccp1 with myelinated axons(NF200) but not compact myelin (MBP) C)They found ccp1 in motor neurons( CHAT) in the ventral horn of spinal cord .
Literature Characterization
This study investigated five mutations introduced into the TEV protease enzyme to improve its solubility. Following the mutations, a technique involving centrifugation was used to evaluate the solubility of each variant. The variants were concentrated ,samples were collected at specific time points, and then the protein concentrations were measured to assess their solubility.
Figure A shows how the concentration of wild-type TEV protein compares to the L56V variant. A plateau in the absorption reading indicates the point at which the protein precipitates out of solution (limited solubility). Introducing mutations L56V and S135G significantly increased TEV solubility (Figure B), allowing them to reach maximum concentrations of 6.2 mg/mL and 5.77 mg/mL, respectively. Since the remaining variants (K45F, Q58F, E106G) displayed similar solubility to the wild-type TEV, they were not investigated further.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 310