Influences of amino acid features of glutathioneS-transferase fusion proteins on their solubility
| Autor: | Mihoko Nagano, Hisashi Koga, Makoto Kawai, Kiyo Shimada |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
DNA
Complementary Recombinant Fusion Proteins Proteomics Biochemistry law.invention Mice chemistry.chemical_compound law Complementary DNA Animals Humans Amino Acid Sequence Amino Acids Antigens Molecular Biology Glutathione Transferase chemistry.chemical_classification biology Decision Trees Glutathione Fusion protein Molecular biology Amino acid Molecular Weight Enzyme Glutathione S-transferase Solubility chemistry biology.protein Recombinant DNA |
| Zdroj: | PROTEOMICS. 5:3859-3863 |
| ISSN: | 1615-9861 1615-9853 |
| DOI: | 10.1002/pmic.200402085 |
| Popis: | We have previously described our strategy for high-throughput (HT) production of recombinant antigens for anti-mKIAA antibody generation, which involves using shotgun fragments generated during entire sequencing of mKIAA cDNAs. We applied this strategy to 1628 mouse KIAA (mKIAA) cDNA fragments, and 84.2% of the GST-mKIAA fusion proteins were successfully purified. The solubility of the proteins was predicted by a small-scale bacterial culture, and a large-scale culture was then performed according to the expected results. Among them, 43.8% of the proteins were purified as a soluble form and 56.2% as an insoluble form. The average yield of the soluble proteins was 0.15 nmol/mL of bacterial culture, and that of the insoluble proteins was 0.55 nmol/mL Statistical analysis of the data revealed a significant correlation between amino acid features of the recombinant proteins and their solubility. To achieve the most effective and feasible protein expression, we constructed a decision tree in which the analyzed data were reflected. The information described here may provide practical guidelines for HT production of recombinant proteins. |
| Databáze: | OpenAIRE |
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