Cloning and Characterization of a cDNA Encoding Aspartate Aminotransferase-P1 from Lupinus angustifolius Root Tips
| Autor: | Brett D. Reddington, Kevin J. F. Farnden, Christopher S. Winefield, William T. Jones, Paul H. S. Reynolds |
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| Rok vydání: | 1994 |
| Předmět: |
clone (Java method)
endocrine system diseases Physiology Phagemid Molecular Sequence Data Aspartate transaminase Saccharomyces cerevisiae Plant Science Biology Molecular cloning Complementary DNA Escherichia coli Genetics Animals Amino Acid Sequence Aspartate Aminotransferases Cloning Molecular Expression vector Base Sequence Sequence Homology Amino Acid cDNA library Genetic Complementation Test nutritional and metabolic diseases DNA Plants Molecular biology Recombinant Proteins Complementation Biochemistry Mutation biology.protein Electrophoresis Polyacrylamide Gel hormones hormone substitutes and hormone antagonists Research Article |
| Zdroj: | Plant Physiology. 104:417-423 |
| ISSN: | 1532-2548 0032-0889 |
| Popis: | A root tip cDNA library, constructed in the [lambda] Zap II expression vector, was immunoscreened with a monoclonal antibody raised against aspartate aminotransferase-P1 from Lupinus angustifolius L. var Uniharvest. One 1452-base pair clone was isolated. The encoded protein sequence had high homology to both plant and animal aspartate aminotransferase sequences. The clone was converted to the phagemid form and expressed in Escherichia coli. The expressed protein was enzymically active and could be immuno-complexed with aspartate aminotransferase-P1-specific antibodies. The complete aspartate aminotransferase-P1 cDNA was cloned into the yeast expression vector pEMBL-yex4 and transformed into Saccharomyces cerevisiae strain BRSCS6, which possesses a mutated aspartate aminotransferase gene (the asp5 mutation). Complementation of the mutation was achieved using this construct. |
| Databáze: | OpenAIRE |
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