Recombinant expression method of human cryptochrome protein I (hcry1) and its application in preparation of radiotherapy protective agent
A cryptochrome and protective agent technology, which is applied to the recombinant expression of human cryptochrome protein I (hCRY1) and its application in the preparation of radiotherapy protective agents, can solve the problems of lack of application prospects, high cost of peptide synthesis, and lack of literature Report on animal CRY1 protein prokaryotic expression and purification methods and other issues
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[0029] The following description of the present invention is based on the examples to further illustrate the present invention, the present invention is not limited to the following examples. For the embodiment of the method of the present invention as follows:
[0030] 1. Construction of prokaryotic expression plasmid pET28a(+)-hCRY1:
[0031] Using the hCRY1 eukaryotic expression plasmid pcDNA3.1-hCRY1 stored in the laboratory as a template, design specific primers according to the sequence of the hCRY1 coding region: the upstream primer is hCRY1FW (5'-ATACTCGAGGCTAAGCCTTCC-3'), and the downstream primer is hCRY1RV (5' -CGCGGATCCTACGTTATACT-3') (both upstream and downstream primers were synthesized by Beijing Aoke Dingsheng Biotechnology Co., Ltd.). PCR amplifies the required target fragment (total system 25uL, 94°C pre-denaturation for 3min, 32 cycles: 94°C for 30s, 55°C for 30s, 72°C for 2min, and finally 72°C for 10 minutes). Use restriction endonucleases XhoI and BamlI...
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