Oncolytic virus with improved safety and anticancer effects
An oncolytic virus, vaccinia virus technology, applied in the direction of anti-tumor drugs, resistance to vector-borne diseases, viruses, etc., can solve the controversial problem of additional cytotoxic effect, and achieve the effect of enhancing anti-tumor effect
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Embodiment 1
[0118] The preparation of the vaccinia virus of embodiment 1 mutation
Embodiment 11
[0119] Example 1.1 Construction of Shuttle Plasmid Vector
[0120] The shuttle plasmid vector used was a shuttle generated by synthesizing the type 1 HSVTK gene (pSE / L promoter) and firefly luciferase reporter (p7.5 promoter) gene and recombining these genes into the pUC57amp+ plasmid (Genewiz, USA) Plasmid vector.
Embodiment 12
[0121] Construction of the vaccinia virus (C1, C3, C40, C45, C52, and C57) of the mutation of embodiment 1.2
[0122] figure 1 The steps for making the mutant vaccinia virus are shown. Specifically, first, in order to obtain the recombinant virus, HeLa cells (ATCC) were cultured at 4×10 per well 5 The cells were seeded in 6-well plates, and then supplemented with EMEM medium containing 10% fetal bovine serum. The Wyeth wild-type vaccinia virus (New York City Department of Health strain, WR-1536, ATCC) was used for treatment at an MOI of 0.05. After 2 hours, replace the medium with EMEM medium containing 2% fetal bovine serum and use Xfect TM Polymer (Clonetech631317, USA) delivered 4 μg of the shuttle plasmid vector (constructed as in Example 1.1) into the cells. After culturing for 4 hours, the culture medium was replaced with EMEM medium containing 2% fetal calf serum, and culture was continued for 72 hours. The recombinant vaccinia virus containing HSV1-TK gene was obt...
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