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Rana wuchuanensis antioxidant peptide wuchuanin-G, gene of coded sequence thereof and application of rana wuchuanensis antioxidant peptide wuchuanin-G
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A technology of antioxidant peptides and coding genes, applied in the field of molecular genetics, to achieve the effect of improving antioxidant effect, small molecular weight, and strong antioxidant activity
Inactive Publication Date: 2014-11-19
GUIZHOU NORMAL UNIVERSITY
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At present, there is no report on the research on antioxidant peptides derived from Wuchuan stink frog
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Embodiment 1
[0021] Example 1 of the present invention: Cloning of the wuchuanin-G gene of the Wuchuan stink frog antioxidant peptide.
[0022] 1) Extraction of total RNA from Wuchuan stink frog skin:
[0023] ①Take 300 mg of Wuchuan stink frog skin tissue, put it into a mortar and add liquid nitrogen to grind it into powder, transfer it to an EP tube, add 1 ml of total RNA extraction buffer (Trizol, a product of Invitrogen, USA), and mix well. Then centrifuge at 12000 rpm for 10 min at 4°C.
[0024] ② Centrifuge to get the supernatant, add 0.2 ml chloroform solution, mix vigorously, let stand at room temperature for 10 minutes, then centrifuge at 4°C, 12000 rpm for 10 minutes, discard the precipitate.
[0025] ③ Add an equal volume of isopropanol to the supernatant, place at room temperature for 10 minutes, centrifuge at 4°C, 12,000 rpm for 10 minutes, collect the precipitate and wash it once with 75% (V / V) ethanol, dry it, and the precipitate at the bottom of the tube is ready for use. ...
Embodiment 2
[0038] Example 2 of the present invention: Chemical synthesis of the antioxidant peptide wuchuanin-G from Wuchuan stink frog.
[0039] Ⅰ. The chemical synthesis method of the antioxidant peptide wuchuanin-G of Wuchuan stink frog: According to the amino acid sequence of the mature peptide deduced from the gene, its complete sequence was synthesized with an automatic peptide synthesizer (433A, Applied Biosystems), and desalted by HPLC reversed-phase column chromatography .
[0040] Ⅱ. Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).
[0041] Ⅲ. Purified wuchuanin-G, an antioxidant peptide of Wuchuan stink frog, was identified by high performance liquid chromatography (HPLC). The molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), and the isoelectric point was determined by isoelectric focusing electrophoresis. , Determining the ami...
Embodiment 3
[0044] Example 3 of the present invention: the application of Wuchuan stink frog antioxidant peptide wuchuanin-G in the preparation of antioxidant drugs.
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Abstract
The invention discloses a rana wuchuanensis antioxidant peptide wuchuanin-G which has a protein with an amino acid sequence shown as SEQ IDNO:1 in a sequence table or a protein which is obtained through substituting, deleting or adding more than one amino acid residue of an amino acid residue sequence shown as SEQIDNO:2, has the same activity as the amino acid residue sequence shown as the SEQIDNO:2 and is derived from the SEQIDNO:2. The rana wuchuanensis antioxidant peptide wuchuanin-G provided by the invention has the characteristics of very strong antioxidant activity, small molecular weight, simple structure, low hemolytic activity and simple preparation method; and the antioxidant effect of an antioxidant drug or cosmetic additive can be improved if the rana wuchuanensis antioxidant peptide wuchuanin-G is applied to the antioxidant drug or cosmetic additive. The gene is an endogenous gene of the rana wuchuanensis, so that the excess expression of the gene cannot affect the safety, and the rana wuchuanensis antioxidant peptide wuchuanin-G can be widely applied to the production of the drug or cosmetic additive.
Description
technical field [0001] The invention relates to the field of molecular genetics, in particular to the gene and application of an antioxidant peptide wuchuanin-G of Wuchuan stink frog and its coding sequence. Background technique [0002] Any atom or group of atoms that contains an unpaired electron is called a free radical. It is very important for all biological organisms to be able to effectively remove toxic active substances from the living environment or their own metabolites. For aerobic respiration organisms, the formation of free radicals, especially active oxygen radicals such as superoxide anion radicals, hydrogen peroxide radicals, etc., is an inevitable by-product in the metabolic process of aerobic respiration. Under normal physiological conditions, mitochondria complete In the process of aerobic respiration, 1-2% of oxygen will be converted into superoxide anion free radicals, and a series of extremely destructive oxidative free radicals such as hydroxyl free ...
Claims
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