Liver cancer superantigen immunogene therapy agent
A gene therapy and superantigen technology, applied in the field of genetic engineering, can solve the problems of unsuccessful research reports, late start, and lag in gene therapy for liver cancer, and achieve the effects of preventing recurrence, enhancing curative effect, and treating metastatic lesions.
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[0061] Embodiment (concrete operation and application):
[0062] (1) Construction of therapeutic target genes SEA and B7.1 and their plasmids with AFP promoter and enhancer as regulatory elements
[0063] 1. Select AFP high-secreting liver cancer cells, isolate the total DNA of the cell genome, design specific primers, and amplify the sequence of the AFP promoter and cis-enhancer on chromosome 4 by PCR, and use CAT (chloramphenicol to acetyl base enzyme) reporter system to detect regulatory activity and ability in CHO cells;
[0064] 2. Nested PCR site-directed mutation of H81, H187, H225, D227 and other sites of SEA, using single-point or combined mutation, inserting the mutant product into the pCDNA3.1 vector, transfecting CHO cells, and detecting the combination of the expressed product and MHCII molecules Ability and immune activation status;
[0065] 3. Through computer software two-dimensional and three-dimensional structure-assisted analysis of linear and spatial stru...
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