Method for preparing dual-regulated oncolytic adenovirus for expressed superantigen gene of targeted prostate tumor
An oncolytic adenovirus and prostate tumor technology, which is applied in the field of anti-prostate tumor expression superantigen gene, double-regulated oncolytic adenovirus preparation, can solve the weakened effect of the virus on tumor cells, fever, and local injection reactions , flu-like symptoms, leukopenia, thrombocytopenia, and other problems, to achieve a strong anti-tumor immune response, improve the effect of killing tumors, and avoid toxic and side effects
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[0026] 1. Reagents needed for experiment:
[0027] The viral vectors pSG502 and HEK293 cells were purchased from Shanghai Hepatobiliary Surgery Hospital Virus and Gene Therapy Center. The virus backbone plasmid PPE3-ccdb-SEA carrying the SEA gene is kept in this room.
[0028] pUC57 cloning vector Sangon
[0029] Taq enzyme shenergy gaming company
[0030] DNA Ligase SolutionI TakaRa Corporation
[0031] Proteinase K, RNase PROMEGA company
[0032] Fetal bovine serum, calf serum, DMEM GIBCO BRL company
[0033] Agarose, ethidium bromide GENE
[0034] Agar powder, tryptone, yeast extract UNIPATH
[0035] Gel recovery kit QIAGEN company
[0036] PCR product recovery kit QIAGEN company
[0037] Plasmid DNA preparation kit QIAGEN company
[0038] Viral DNA extraction kit QIAGEN company
[0039] LipofectAmine2000 kit GIBCO BRL
[0040] 2. Cloning of prostate-specific promoter:
[0041] According to the sequence (U37672.1) obtained by NCBI Nucleotide in the United States, design and amplify the upstre...
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