Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli
| Autor: | Hui-Yuan Wu, Ruixue Wang |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
lcsh:GE1-350
0301 basic medicine Polyglutamate biology Chemistry GroES medicine.disease_cause biology.organism_classification GroEL law.invention Chaperonin 03 medical and health sciences Oleispira antarctica 030104 developmental biology 0302 clinical medicine Biochemistry law medicine Recombinant DNA Escherichia coli lcsh:Environmental sciences 030217 neurology & neurosurgery Bacteria |
| Zdroj: | E3S Web of Conferences, Vol 185, p 04050 (2020) |
| ISSN: | 2267-1242 |
| DOI: | 10.1051/e3sconf/202018504050 |
| Popis: | Cytosolic carboxypeptidase 6 (CCP6) is a member of cytosolic carboxypeptidase (CCP) family that catalyze the removal of polyglutamate side chains from protein substrates. Biochemical and biophysical characterization of CCPs requires large quantities of purified proteins. However, no method describing the expression of any mammalian CCP family member from bacteria has been published to our best knowledge. After considerable efforts to improve the solubility of mammalian CCPs expressed in bacteria, including the optimization of induction temperature and by using different receptive cells, we were able to get less expression of mouse CCP6 in soluble fraction of bacterial lysates. We report in this article, the bacterial expression of CCP6 using Arctic Express (DE3) competent cells that co-express the chaperonin system GroEL and GroES from Oleispira antarctica. However, to achieve a large number of soluble target proteins, the expression conditions still need to be further optimized. |
| Databáze: | OpenAIRE |
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