Antiviral fusion protein and application thereof
A fusion protein, antiviral technology, applied in antiviral agents, peptide/protein components, hybrid peptides, etc., can solve the problems of high cost, difficult for patients to adhere to lifelong medication, and large side effects, and achieve good practical prospects.
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[0048] 1. Protein preparation method
[0049] 1. Gene cloning to construct target protein expression vector
[0050] 1.1 Construction of pET-15b-PEP-1 plasmid
[0051] Two oligonucleotide chains of 21 amino acids of PEP1 (KETWWETWWTEWSQPKKKRKV) were chemically synthesized, and NdeI (CATATG) and XhoI (CTCGAG) restriction site sequences were added at both ends according to the characteristics of the selected pET-15b expression vector. The sequence is as follows:
[0052] Sequence 1:
[0053] 5'-NdelTATGAAAGAAACCTGGTGGGAAACCTGGTGGACCGAATGGTCTCAGCCGAAAAAAAAAACGTAAAGTGC-3'(68bp)
[0054] Sequence 2:
[0055] 3'-ACTTTCTTTGGACCACCCTTTTGGACCACCTGGCTTACCAGAGTCGGCTTTTTTTTTGCATTTCACGAGCT-5'Xhol(70bp)
[0056] Take out the two tubes of dry powder fragments, centrifuge at 3000rpm for 1min, mix the two fragments equimolarly with STE buffer (pH8.0), incubate at 94°C for 7min, turn off the water bath, let it slowly cool to room temperature for renaturation, and form the PEP-1 encoded DN...
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