The expression of a synthetic rainbow trout metallothionein gene in E. coli
| ISSN: |
0167-4781
|
|---|---|
| Keywords: |
(Rainbow trout) ; Cd stabilisation ; E. coli expression ; Enzyme purification ; Immunochemical characterization ; Metallothionein ; Recombinant protein
|
| Source: |
Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
|
| Topics: |
Biology
Chemistry and Pharmacology
Medicine
Physics
|
| Type of Medium: |
Electronic Resource
|
| URL: |
| _version_ | 1798292540669034496 |
|---|---|
| autor | Kille, P. Stephens, P. Cryer, A. Kay, J. |
| autorsonst | Kille, P. Stephens, P. Cryer, A. Kay, J. |
| book_url | http://linkinghub.elsevier.com/retrieve/pii/0167-4781(90)90054-6 |
| datenlieferant | nat_lic_papers |
| fussnote | A synthetic gene for rainbow trout metallothionein was constructed and inserted into a dual origin plasmid where expression was induced by a temperature shift in a proteinase-deficient strain of Escherichia coli. The recombinant protein was purified to homogeneity, and a partial amino acid sequence was determined to confirm its identity. Its immunochemical characteristics were similar to those of native metallothionein from rainbow trout. The amounts of recombinant metallothionein produced were quantified in soluble cell extracts by ELISA. Low concentrations were detected when growth was performed either in L-broth or defined (GMM-II) medium. Supplementation of the medium with zinc or copper had no effect on the amount of metallothionein produced. By contrast, when cadmium was included in either L-broth or GMM-II medium, much higher concentrations of the protein within the cells (approx. 13 μg/mg soluble cell protein) were detected. This stabilisation of the protein by metal reconstitution in vivo is considered in relation to the selective uptake / exclusion of metals by the cells and its significance for the scavenging of certain precious or toxic heavy metals is discussed. |
| hauptsatz | hsatz_simple |
| identnr | NLZ186816723 |
| issn | 0167-4781 |
| journal_name | Biochimica et Biophysica Acta (BBA)/Gene Structure and Expression |
| materialart | 1 |
| package_name | Elsevier |
| publikationsort | Amsterdam |
| publisher | Elsevier |
| reference | 1048 (1990), S. 178-186 |
| schlagwort | (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein |
| search_space | articles |
| shingle_author_1 | Kille, P. Stephens, P. Cryer, A. Kay, J. |
| shingle_author_2 | Kille, P. Stephens, P. Cryer, A. Kay, J. |
| shingle_author_3 | Kille, P. Stephens, P. Cryer, A. Kay, J. |
| shingle_author_4 | Kille, P. Stephens, P. Cryer, A. Kay, J. |
| shingle_catch_all_1 | Kille, P. Stephens, P. Cryer, A. Kay, J. The expression of a synthetic rainbow trout metallothionein gene in E. coli (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein 0167-4781 01674781 Elsevier |
| shingle_catch_all_2 | Kille, P. Stephens, P. Cryer, A. Kay, J. The expression of a synthetic rainbow trout metallothionein gene in E. coli (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein 0167-4781 01674781 Elsevier |
| shingle_catch_all_3 | Kille, P. Stephens, P. Cryer, A. Kay, J. The expression of a synthetic rainbow trout metallothionein gene in E. coli (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein 0167-4781 01674781 Elsevier |
| shingle_catch_all_4 | Kille, P. Stephens, P. Cryer, A. Kay, J. The expression of a synthetic rainbow trout metallothionein gene in E. coli (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein (Rainbow trout) Cd stabilisation E. coli expression Enzyme purification Immunochemical characterization Metallothionein Recombinant protein 0167-4781 01674781 Elsevier |
| shingle_title_1 | The expression of a synthetic rainbow trout metallothionein gene in E. coli |
| shingle_title_2 | The expression of a synthetic rainbow trout metallothionein gene in E. coli |
| shingle_title_3 | The expression of a synthetic rainbow trout metallothionein gene in E. coli |
| shingle_title_4 | The expression of a synthetic rainbow trout metallothionein gene in E. coli |
| sigel_instance_filter | dkfz geomar wilbert ipn albert fhp |
| source_archive | Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 |
| timestamp | 2024-05-06T08:50:08.941Z |
| titel | The expression of a synthetic rainbow trout metallothionein gene in E. coli |
| titel_suche | The expression of a synthetic rainbow trout metallothionein gene in E. coli A synthetic gene for rainbow trout metallothionein was constructed and inserted into a dual origin plasmid where expression was induced by a temperature shift in a proteinase-deficient strain of Escherichia coli. The recombinant protein was purified to homogeneity, and a partial amino acid sequence was determined to confirm its identity. Its immunochemical characteristics were similar to those of native metallothionein from rainbow trout. The amounts of recombinant metallothionein produced were quantified in soluble cell extracts by ELISA. Low concentrations were detected when growth was performed either in L-broth or defined (GMM-II) medium. Supplementation of the medium with zinc or copper had no effect on the amount of metallothionein produced. By contrast, when cadmium was included in either L-broth or GMM-II medium, much higher concentrations of the protein within the cells (approx. 13 μg/mg soluble cell protein) were detected. This stabilisation of the protein by metal reconstitution in vivo is considered in relation to the selective uptake / exclusion of metals by the cells and its significance for the scavenging of certain precious or toxic heavy metals is discussed. |
| topic | W V WW-YZ U |
| uid | nat_lic_papers_NLZ186816723 |