A recombinant collagen product for skin photodamage repair
A technology of recombining collagen and photodamage, applied in the field of dermatology, can solve the problems of easy inactivation of collagen, complicated processing process, difficult quality control, etc., and achieve the effect of good water solubility, good biocompatibility, and water retention.
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Embodiment 1
[0035] The preparation of embodiment 1 recombinant collagen
[0036] 1. Determine the amino acid sequence comprising recombinant collagen as shown below:
[0037] MHHHHHHADEQEEKAKVRTELIQELAQGLGGFEKKNFPTLGDEDLDHTYMTKLLTYLQEREQAENSWRKRLLKGIQDHALDLVPRGSPGLPGPRGEQGPTGPTGPAGPRGLQGLQGLQGERGEQGPTGPAGPRGLQGERGEQGPTGLAGKAGEAGAKGETGPAGPQGPRGEQGPQGLPGKDGEAGAQGRPGKRGKQGQKGEKGEPGTQGAKGDRGETGPVGPRGERGEAGPAGKDGERGFPGERGVEGQNGQDGLPGKDGKDGQNGKDGLPGKDGKDGQNGKDGLPGKDGKDGQDGKDGLPGKDGKDGLPGKDGKDGQPGKPGKYGPPGPPGPPGPPGPPGPPGPPGPPGPPGPP。
[0038] 2. Construction of recombinant collagen expression strains
[0039] Synthesize the nucleotide sequence encoding the amino acid sequence described in 1 above, construct the expression plasmid containing the nucleotide sequence, and confirm the successful synthesis of the plasmid by DNA sequencing; transform the plasmid into Escherichia coli BL21-DE3 strain, and transform the successfully transformed positive clone After adding glycerin to the strain, store i...
Embodiment 2
[0043] The detection of embodiment 2 recombinant collagen
[0044] 1. Circular Dichroism Detection
[0045] The recombinant collagen prepared in Example 1 was dissolved in PBS buffer solution (pH 7.0), sodium acetate buffer solution (pH 3.0) and ethylene glycol, respectively, with a final concentration of 1 mg / mL. The circular dichroism spectrometer was used to scan the spectrum of the above-mentioned recombinant collagen solution. The scanning wavelength was 210-260nm, the scanning temperature was 4°C, the wavelength step was 0.5nm, and the average time was 0.5 seconds. The CD spectrum was the average value of three scans.
[0046] The result is as figure 1 As shown, the recombinant collagen prepared in Example 1 of the present invention has a positive peak at around 220 nm, which conforms to the triple helical structure characteristic of collagen, and is a recombinant collagen with a triple helix structure.
[0047] 2. Cytotoxicity experiment
[0048] HeLa cells grown com...
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