| Autor: | Paul A. Millner, Murphy Ga, Wise A, Carr Th, Thomas Pg |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
biology
G protein Plant Science General Medicine medicine.disease_cause biology.organism_classification Molecular biology law.invention chemistry.chemical_compound Biochemistry chemistry Affinity chromatography law Arabidopsis Gene expression Genetics Recombinant DNA medicine Agarose PMSF Agronomy and Crop Science Escherichia coli |
| Zdroj: | Plant Molecular Biology. 33:723-728 |
| ISSN: | 0167-4412 |
| DOI: | 10.1023/a:1005732423622 |
| Popis: | The Arabidopsis Gα subunit, GPα1, was expressedwithin Escherichia coli by co-transformation with the expressionvector and the dnaY gene which encodes tRNAArg AGA/AGG. Isolation of the recombinant GPα1 in a highly pureform could be achieved by a combination of anion exchange and dyeaffinity chromatography or by a single step affinity procedure viachromatography on 4-amino-anilido-GTP agarose. The recombinant proteinyielded by both procedures was highly active and bound GTPγS withan apparent Kd in the nM range. GTPγS binding wasstimulated two-fold in the presence of Zn2+ compared with that inthe presence of Mg2+, Mn2+ or Ca2+.Abbreviations: 4aaGTP, 4-amino-anilido-GTP; GTPγS,guanosine- 5′-(3-O-thiotriphosphate), PMSF,phenylmethylsulphonyl fluoride; PVDF, polvinylidene fluoride;rGPα1, recombinant GPα1 |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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