Polypeptide CJ130 with anti-HIV (human immunodeficiency virus)-1 activity and application thereof
A technology for HIV-1 and HIV-1B, which is applied to the anti-HIV-1 active polypeptide CJ130 and its application field, which can solve the problems of toxic side effects and the inability to completely remove the virus.
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Embodiment 1
[0021] Embodiment 1, the preparation of polypeptide
[0022] The inventors of the present invention have designed polypeptide CJ130, the amino acid sequence of which is as follows:
[0023] Polypeptide CJ130 (SEQ ID NO: 1 of the Sequence Listing): VWSIVIIEYRKILRQRKIDRRRRRRRRRR.
[0024] Artificially synthesized polypeptide CJ130.
Embodiment 2
[0025] Embodiment 2, the toxicity determination of polypeptide
[0026] 1. Dissolve 1 mg of the polypeptide CJ130 prepared in Example 1 in 20 μl of DMSO, and then perform 2-fold gradient dilution with DMSO to obtain polypeptide dilutions containing different concentrations of the polypeptide.
[0027] 2. Take the white transparent 96-well plate and add the polypeptide dilution prepared in step 1 (1 μl per well).
[0028] 3. Suspend MT-2 cells in RPMI1640 medium containing 10% fetal bovine serum by volume to obtain 1.5×10 5 cells / ml of MT-2 cell suspension.
[0029] 4. Take the MT-2 cell suspension obtained in step 3, add it to the 96-well plate (200 μl per well) that completed step 2, centrifuge briefly, and then 37 ° C, 5% CO 2 Incubate for 72 hours.
[0030] 5. After completing step 4, take the 96-well plate and add 20 μl CellTiter- Luminescent Cell Viability Assay (CellTiter- Luminescence method cell viability detection kit), the luminescence signal was measured by W...
Embodiment 3
[0033] Embodiment 3, the anti-HIV-1 activity determination of polypeptide
[0034]1. Dissolve 1 mg of the polypeptide CJ130 prepared in Example 1 in 20 μl of DMSO, and then perform 2-fold gradient dilution with DMSO to obtain polypeptide dilutions containing different concentrations of the polypeptide.
[0035] 2. Take the white transparent 96-well plate, add the polypeptide dilution prepared in step 1 (1 μl per well)
[0036] 3. Suspend MT-2 cells with RPMI1640 culture medium containing 10% fetal bovine serum by volume percentage to obtain 3.0×10 5 cells / ml of MT-2 cell suspension.
[0037] 4. Adjust the virus concentration in the virus solution of pNL4.3 virus with RPMI1640 culture solution containing 10% fetal bovine serum by volume percentage to obtain a virus dilution solution of 400 TCID50 / ml.
[0038] 5. Mix the MT-2 cell suspension obtained in step 3 with the virus dilution obtained in step 4 in equal volumes, then add the mixed solution to the 96-well plate (200 μl ...
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