A heptapeptide anti-aβ protein aggregation and its application and gene encoding the synthetic polypeptide
A β-protein and heptapeptide technology, applied in the field of heptapeptide and its application and the gene encoding the synthetic polypeptide, can solve problems such as lack of specificity, and achieve the effects of improving memory, improving medical conditions, and major social and economic benefits
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Embodiment 1
[0041] Synthesis of Peptide WW-7 by Solid Phase Synthesis of Peptide
[0042] 1. Resin selection
[0043] (1) Using the standard Fomc protocol, starting with 0.0125mmol 2-chlorotrityl chloride resin resin (Tianjin Nankai Synthetic Technology Co., Ltd.), according to the amino acid sequence Trp-Asp-Gln-Trp-Cys-Ile-Trp from the C-terminal to the N According to the sequence characteristics of the terminal, 0.3mol of the first Fmoc protected amino acid was added, DCC and 5% (mass fraction) DMAP were added to the reactor to shake the reaction, and the resin was washed with methylpyrrolidone (NMP) to remove excess protected amino acids.
[0044] (2) Using the standard Fomc scheme, initially select 0.0125mmol Wang resin, according to the sequence characteristics of the amino acid sequence Trp-Asp-Gln-Trp-Cys-Ile-Trp from the C-terminal to the N-terminal, add 0.3mol of the first Fmoc protected amino acid , DCC and 5% (mass fraction) DMAP are added to the reactor for shaking reaction,...
Embodiment 2
[0052] Anti-Aβ42 Protein Aggregation Activity Experiment of Polypeptide WW-7 in Vitro
[0053] 1. Experimental method
[0054] Preparation of culture medium: High glucose medium (DMEM), fetal bovine serum (FBS), L-glutamine were prepared according to the mass ratio of 8.75:1:0.25 respectively; 1wt% double antibodies (penicillin and streptomycin) were added at the same time , 0.1 wt% Hygromycin B and 0.05 wt% Blasticidin S antibiotics.
[0055] Preparation of 0.05mM and 0.5mM polypeptide (WW-7) solutions: Weigh 10.4mg of polypeptide WW-7, dissolve it with 10mL of medium, pass through a 0.22μm filter head, the concentration of the mother solution is 1mM, and then use the medium to dissolve The mother liquor was diluted to the concentration required for the experiment.
[0056] Preparation of 1 mg / mL tetracycline solution: Weigh 20 mg of tetracycline, prepare with 10 mL of PBS buffer, pass through a 0.22 μm filter head, and store at -20°C in the dark for future use.
[0057] F...
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