High level expression of a synthetic gene coding for IgG-binding domain B of Staphylococcal protein A
| Autor: | Kei C O Patent Departm Okazaki, Seiga Itoh, Moriyuki Sato, Akiko Saito, Shinkichi Honda, Masamichi Koike, Tatsunari Nishi |
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| Rok vydání: | 1989 |
| Předmět: |
Molecular Sequence Data
Bioengineering Biology Protein Engineering Biochemistry Fusion gene chemistry.chemical_compound Genes Synthetic Amino Acid Sequence Binding site Staphylococcal Protein A Molecular Biology Chromatography High Pressure Liquid Base Sequence Protein engineering Fusion protein Restriction enzyme Gene Expression Regulation chemistry GATAD2B Immunoglobulin G biology.protein Electrophoresis Polyacrylamide Gel Cyanogen bromide Protein A Biotechnology |
| Zdroj: | "Protein Engineering, Design and Selection". 2:481-487 |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/2.6.481 |
| Popis: | A gene coding for one of the IgG-binding domains of Staphylococcal protein A, designated domain B, was chemically synthesized. This gene was tandemly repeated to give dimeric and tetrameric domain B genes by the use of two restriction enzymes which gave blunt ends. The genes were highly expressed in Escherichia coli to afford a large amount of dimeric and tetrameric domain B proteins. The single domain B protein was efficiently produced as a fusion protein with a salmon growth hormone fragment. The fusion protein was converted to monomeric domain B by cyanogen bromide cleavage. The CD spectra of the monomeric, dimeric and tetrameric domain B proteins were essentially the same as that of native form protein A, showing that their secondary structures were very similar. The dimeric and tetrameric domain B proteins formed precipitates with IgG as protein A. This system permits the efficient production of mutated single and multiple IgG-binding domains which can be used to study structural changes and protein A-immunoglobulin interactions. |
| Databáze: | OpenAIRE |
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