Expression of a Bifunctional Chimeric Protein A-Vargula hilgendorfii Luciferase in Mammalian Cells
| Autor: | T Hara, Genji Kawano, Eiji Suzuki, Hiroshi Ueda, J Kazami, Yumi Maeda, Teruyuki Nagamune |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Recombinant Fusion Proteins
Molecular Sequence Data Gene Expression Vargula hilgendorfii CHO Cells General Biochemistry Genetics and Molecular Biology Cell Line Cricetinae Animals Humans Luciferase Amino Acid Sequence Luciferases Staphylococcal Protein A Peptide sequence DNA Primers Immunoassay Rous sarcoma virus Base Sequence biology Chinese hamster ovary cell Structural gene biology.organism_classification Molecular biology Fusion protein Coleoptera Evaluation Studies as Topic Immunoglobulin G Luminescent Measurements biology.protein Protein A Plasmids Biotechnology |
| Zdroj: | BioTechniques. 20:116-121 |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/96201rr01 |
| Popis: | We have designed and constructed a novel chimeric protein that consisted of a single domain of protein A and luciferase derived from sea-firefly Vargula hilgendorfii with the goal of obtaining a heterofunctional immunological tool. The structural gene of luciferase was fused to the 3′ terminus of the D domain gene of protein A with/without a short linker of five amino acids. The resulting constructs under the transcriptional regulation of the Rous sarcoma virus (RSV) promoter, were expressed transiently in simian COS-1 and stably in Chinese hamster ovary (CHO) cells. The properties of the resultant chimeric protein were characterized. The results indicated that the dual properties of the chimeric protein could be retained only after the introduction of a linker of (Gly)4 Ser between the two conjugated moieties. Moreover, the chimeric protein was found to retain at least 50% of the specific activity as compared with the non-fused luciferase. The future prospect of the usage of this chimeric protein in the field of diagnostics was further evaluated by performing bioluminescent immunoassays. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|