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Artigo em Inglês | WPRIM | ID: wpr-263230

RESUMO

v-Src is a non-receptor protein tyrosine kinase involved in many signal transduction pathways and closely related to the activation and development of cancers. We present here the expression, purification, and bioactivity of a GST (glutathione S-transferase)-fused v-Src from a bacterial expression system. Different culture conditions were examined in an isopropyl beta-D-thiogalactopyranoside (IPTG)-regulated expression, and the fused protein was purified using GSH (glutathione) affinity chromatography. ELISA (enzyme-linked immunosorbent assay) was employed to determine the phosphorylation kinase activity of the GST-fused v-Src. This strategy seems to be more promising than the insect cell system or other eukaryotic systems employed in earlier Src expression.


Assuntos
Proteínas de Bactérias , Química , Genética , Glutationa Transferase , Genética , Proteína Oncogênica pp60(v-src) , Química , Genética , Engenharia de Proteínas , Métodos , Proteínas Recombinantes de Fusão , Química , Saccharomyces cerevisiae , Genética , Metabolismo
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