Application of human tpt1/tctp gene in preparation of antineoplastic drugs
A gene and drug technology, applied in the field of preparation of anti-tumor drugs, can solve the problems that there are no related reports
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Embodiment 1
[0016] Example 1 Construction of TPT1 lentiviral vector and establishment of stably transfected cell lines
[0017] 1. siRNA target design
[0018] The nucleotide sequence of human TPT1 (NM_001286272.1) was queried at the National Center for Biological Information (NCBI), and 12 pairs of effective siRNA targets were designed for the coding sequence region using DSIR software.
[0019]
[0020] 2. Plasmid construction of pLVX-shRNA2-Puro-TPT1 and negative control pLVX-shRNA2-Puro-NC
[0021] According to the above siRNA sequence (taking SEQ ID NO: 1, 2, 3 as an example), design corresponding complementary single-stranded DNA, and add BamHI and EcoRI restriction sites at both ends.
[0022]
[0023] Annealing: Anneal the upstream and downstream fragments of single-stranded shRNA to form double-stranded DNA fragments, which are then ligated to the digested vector.
[0024] Annealing reaction system:
[0025]
[0026] Amplification program: 95°C, 5 min; cool at room te...
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