Sheng wu hua hsüeh yü sheng wu wu li hsüeh pao, 2002-03, Vol.34 (2), p.248
2002
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- Autor(en) / Beteiligte
- Titel
- Functional di-domain of neuronal growth inhibitory factor
- Ist Teil von
- Sheng wu hua hsüeh yü sheng wu wu li hsüeh pao, 2002-03, Vol.34 (2), p.248
- Ort / Verlag
- China
- Erscheinungsjahr
- 2002
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Neuronal growth inhibitory factor (GIF), known also as metallothionein-III (MT-III), was the first validated to be capable of inhibiting growth of neuronal cells in nervous system, its beta-domain being functional. GIF functional di-domain (GIFbeta- beta) was constructed to study the structure and function of GIF. N terminal beta-domain and C terminal beta-domain cDNAs were amplified by PCR, inserted into vector pGEX-4T-1 and expressed in Escherichia coli, as carboxyl terminal extension of glutathione-S-transferase (GST), by IPTG induction. After digestion by thrombin, the fusion protein was isolated by passing through a glutathione-Sepharose 4B affinity chromatography column and was purified by gel fit ration on Sephacryl-S100. About 60 mg protein per liter of bacterial cell culture was achieved. The results of SDS-PAGE, amino acid composition, molecular mass, the ratio of metal/protein and sulfhydryl group/protein showed that the purified protein was the GIFbeta- beta. Circular dichroism (CD) spectroscopy s
- Sprache
- Chinesisch
- Identifikatoren
- ISSN: 0582-9879Titel-ID: cdi_pubmed_primary_12007006
- Format
- –
- Schlagworte
- Animals, Binding Sites - genetics, Cell Division - drug effects, Cells, Cultured, Circular Dichroism, DNA, Complementary - genetics, Electrophoresis, Polyacrylamide Gel, Escherichia coli - genetics, Female, Mice, Nerve Tissue Proteins - chemistry, Nerve Tissue Proteins - genetics, Nerve Tissue Proteins - pharmacology, Neurons - cytology, Neurons - drug effects, Neurons - metabolism, Peptide Fragments - chemistry, Peptide Fragments - genetics, Peptide Fragments - pharmacology, Plasmids - genetics, Recombinant Proteins - chemistry, Recombinant Proteins - metabolism, Recombinant Proteins - pharmacology