Construction method for prokaryotic secretory expression vector of protein transduction domain-Apoptin (PTD-Apoptin) fusion protein and application of prokaryotic secretory expression vector
The invention relates to a construction method for a prokaryotic secretory expression vector of a protein transduction domain-Apoptin (PTD-Apoptin) fusion protein and the application of the prokaryotic secretory expression vector. The construction method comprises the following steps of: connecting...
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| creator | CUI JIAN BEN SONGBIN CHEN CHANGLAN LIU XUEMEI LI QIJIU |
| description | The invention relates to a construction method for a prokaryotic secretory expression vector of a protein transduction domain-Apoptin (PTD-Apoptin) fusion protein and the application of the prokaryotic secretory expression vector. The construction method comprises the following steps of: connecting a PTD-Apoptin sequence onto a pET22b(+) secretory expression vector to construct the prokaryotic secretory expression vector pET22b(+)-PTD-Apoptin; secreting an expressed interest protein into the periplasmic space of escherichia coli through isopropyl thiogalactoside (IPTG) induction; extracting the protein of the periplasmic space by an osmose process; performing ultrasonication on remaining cellular components; then, separating supernatant and depositing; and performing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Co-incubation is performed on the PTD-Apoptin fusion protein in the periplasmic space and a gastric cancer cell-823, and the inhibitory actionof the fusion protein on |
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The construction method comprises the following steps of: connecting a PTD-Apoptin sequence onto a pET22b(+) secretory expression vector to construct the prokaryotic secretory expression vector pET22b(+)-PTD-Apoptin; secreting an expressed interest protein into the periplasmic space of escherichia coli through isopropyl thiogalactoside (IPTG) induction; extracting the protein of the periplasmic space by an osmose process; performing ultrasonication on remaining cellular components; then, separating supernatant and depositing; and performing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. 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| language | chi ; eng |
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| subjects | BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS THEREOF CULTURE MEDIA ENZYMOLOGY HUMAN NECESSITIES HYGIENE MEDICAL OR VETERINARY SCIENCE METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS SPIRITS VINEGAR WINE |
| title | Construction method for prokaryotic secretory expression vector of protein transduction domain-Apoptin (PTD-Apoptin) fusion protein and application of prokaryotic secretory expression vector |
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