Polypeptides of anti-measles virus and human immunodeficiency virus and use
A technology of immunodeficiency virus and measles virus, which is applied in the field of anti-measles virus and human immunodeficiency virus anti-virus polypeptide of East Asian scorpion, and the field of polypeptide with high-efficiency anti-virus function, which can solve the instability and preventive effect of its own genetic material Reduce the problem of easy mutation and other problems, and achieve the effect of good water solubility, good inhibitory effect and no side effects
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Embodiment 1
[0025]Example 1: Preparation of a scorpion venom antiviral polypeptide. Proceed as follows:
[0026] A: Extraction of total RNA from scorpion venom glands (Trizol LS one-step method: Trizol LS was purchased from Invitrogen, USA)
[0027] ① Grind 500mg of scorpion glands into fine powder in liquid nitrogen, add 10ml Trizol LS to mix, and place at room temperature (20-25°C, the same below) for 5 minutes; Minutes, centrifuge at 12,000g for 15 minutes at 4°C; ③Add 1 volume of isopropanol to the water phase, place at room temperature for 10 minutes, and centrifuge at 12,000g for 10 minutes at 4°C to obtain RNA precipitation; ④Wash the precipitate with 5ml of 75% ethanol, and centrifuge at 7,500g for 5 minutes ; ⑤ After the RNA precipitate was dried, it was dissolved in DEPC (diethypyrocarbonate, diethyl pyrocarbonate)-treated sterilized water, and kept at 55-60°C for 10 minutes to completely dissolve the RNA. The whole process was carried out according to the method recommended b...
Embodiment 2
[0075] Example 2: Chemical synthesis of scorpion venom antiviral polypeptide AVP-W1
[0076] A polypeptide AVP-W1 consisting of 13 amino acids was isolated from the venom gland of the East Asian scorpion and was amidated. A high-purity AVP-W1 polypeptide: FVAGVASLVRIKF-NH2 was obtained by solid-phase chemical synthesis.
Embodiment 3
[0077] Example 3: Inhibition of Measles Virus by Antiviral Polypeptide AVP-W1 Drugs
[0078] Material:
[0079] A: Culture of Vero E6 cells
[0080] The African green monkey kidney epithelial cell line (VeroE6) was purchased from China Center for Type Culture Collection.
[0081] Medium: 10% (passage medium) or 2% (maintenance medium) calf serum (FCS, GIBCO-BRL), 56 ° C, 30 minutes heat sterilization; 2mM / mL L-glutamate; 100U / mL Penicillin and 100 μg / mL streptomycin. Culture conditions are: 37°C, 5% CO 2 Incubator cultivation.
[0082] B: virus strain
[0083] The Edmonston measles virus used in this study came from the Wuhan Institute of Virology, Chinese Academy of Sciences (purchased from the American Type Culture Collection).
[0084] Specific operation method:
[0085] Monolayer Vero E6 cells were inoculated in a 96-well plate, 10 per well 4 For cells, add resistant medium to allow the cells to grow and adhere to the wall. The virus was added to the maintenance m...
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