Recombinant Escherichia coli for expressing Caspase-3 recombinant scFv78 and functional verification method of recombinant Escherichia coli
A technology for recombining Escherichia coli and scfv78, applied in the field of molecular biology, to achieve great application value, clear genome and genetic background, and easy transformation
Pending Publication Date: 2020-05-01
NANCHANG UNIV
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Abstract
The invention provides recombinant Escherichia coli for expressing Caspase-3 recombinant scFv78 and a functional verification method of the recombinant Escherichia coli. The method comprises the following steps: firstly, connecting a recombinant single-chain antibody scFv78 encoding gene integrated with a Caspase-3 encoding gene to a pET302 vector through an enzyme digestion enzyme-linked approach, and converting obtained recombinant plasmid into Escherichia coli Nissle1917 so as to obtain a bacterial strain capable of expressing the Caspase-3 recombinant scFv78. The invention adopts the intracellular invasion and aggregation characteristics of the Escherichia coli Nissle1917 and the fixed-point targeting characteristic of the scFv78 on TEM1 positive cells; the human-derived toxin Caspase-3 protein site plays a role specifically, so that the TEM1 positive tumor cells are killed more accurately, and damage to normal cells of an organism is reduced. On the basis, the method utilizes recombinant bacteria to express the human-derived toxin Caspase-3 recombinant single-chain antibody scFv78 protein, and then the protein and MS1-TEM1 cells are co-incubated in an in-vitro environment, sothat the affinity and lethality of the to bacterial strain tumor cells are verified.
Application Domain
BacteriaMicroorganism based processes +3
Technology Topic
Ribosomal RNA GenesMolecular biology +12
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