Anti-tumor DNA vaccine and preparation method thereof
A DNA vaccine and anti-tumor technology, applied in the field of medicine, can solve the problems of weak tumor targeting and DNA vaccine immunogenicity, and achieve the effect of improving immunogenicity, high safety, and strong targeting
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example 1
[0069] 1. Acquisition of mtHsp70 and HSV-tk genes
[0070] (1) Design primers:
[0071] According to the mRNA sequences of HSV-TK (genbank NO.J02224.) and Mycobacterium tuberculosis Hsp70 gene (genbank NO.BX842573), the specific primers for amplifying the full-length cDNA of HSV-TK and mtHsp70 genes were respectively designed by using the primer design software DNA club. The sequence is as follows:
[0072] HSV-TK primers:
[0073] Forward: 5 GCGCCTTGTAGA AGCGCG TA-3' (SEQ NO.1)
[0074] Reverse: 5' CCTTCCGGTATTGTCTCCCTTCC-3' (SEQ NO.2)
[0075] Both the upstream primer and the downstream primer introduce a restriction endonuclease Not I site (framed partial sequence);
[0076] Hsp 70 primer:
[0077] Forward: 5 CGCCTACATGGCTCGTGC-3' (SEQ NO.3)
[0078] R Reverse: 5 TCACTTGGCCTCCCGG-3' (SEQ NO.4)
[0079] Upstream and downstream primers introduce restriction endonuclease Xho I and EcoR I sites (framed partial sequences) respectively;
[0080] Submit the above-des...
example 2
[0147] 1. Acquisition of mt Hsp70 and HSV-tk genes: the same as Example 1.
[0148] 2. Construction of pCMV-mt Hsp70-IRES-tk: same as Example 1.
[0149] 3. Preparation of the DNA vaccine of the present invention: transform pCMV-mtHsp70-IRES-tk into Lactobacillus, and the specific operation is the same as in Example 1.
example 3
[0151] 1. Acquisition of mt Hsp70 and HSV-tk genes: the same as Example 1.
[0152] 2. Construction of pCMV-mt Hsp70-IRES-tk: same as Example 1.
[0153] 3. Preparation of the DNA vaccine of the present invention: transform pCMV-mt Hsp70-IRES-tk into bifidobacteria, and the specific operation is the same as in Example 1.
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