Novel recombinant protein FCN1A-QTY-COIA1 and application thereof
By modifying and designing the FCN1A_XENLA and COIA1_HUMAN proteins, a new recombinant protein FCN1A-QTY-COIA1 was formed, which solved the problems of complex separation and purification of natural proteins and limited effectiveness, and achieved effective treatment of atopic dermatitis and cost reduction.
Patent Information
- Application Number
- CN202510956886.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-11
- Publication Date
- 2025-10-10
AI Technical Summary
Existing natural proteins used in the treatment of atopic dermatitis have problems such as complex separation and purification processes, high costs and limited effects, which make it difficult to meet clinical needs.
The FCN1A_XENLA and COIA1_HUMAN proteins were modified and designed through synthetic biology methods, and some amino acid sequences were mutated and connected with a flexible protein linker to form a new recombinant protein FCN1A-QTY-COIA1, which retains the original function and improves hydrophilicity.
The new recombinant protein FCN1A-QTY-COIA1 significantly improves atopic dermatitis, has good skin repair effects, reduces production costs, and improves biocompatibility and hydrophilicity.
Smart Images
Figure CN120757666A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biomedicine technology, and in particular to a novel recombinant protein FCN1A-QTY-COIA1 and applications thereof. Background Art
[0002] Atopic dermatitis (AD) is a common chronic inflammatory skin disease characterized by intractability, recurrence, and pruritus. Currently, AD treatments include topical medications such as glucocorticoids and calcineurin inhibitors, as well as systemic medications such as oral antihistamines, immunosuppressants, and biologics. However, these treatments have limitations. For example, long-term use of glucocorticoids may lead to side effects such as skin atrophy and capillary dilation, while biologics are expensive and may pose immunogenicity issues.
[0003] Protein solutions or protein-based hydrogels of a certain concentration have good moisturizing properties, can maintain skin moisture, relieve skin dryness and itching symptoms, regulate the physical and chemical properties of the skin, promote the proliferation and differentiation of skin cells, and accelerate skin repair and regeneration. FCN1A_XENLA and COIA1_HUMAN proteins are two natural proteins with good biocompatibility, biodegradability and low immunogenicity, and can effectively promote cell growth, proliferation and differentiation. FCN1A_XENLA protein belongs to the fibrogel protein family (FCN), and COIA1_HUMAN protein belongs to the collagen family. Previous studies have shown that the above two family proteins have certain properties of improving atopic dermatitis, but the effect is not significant.
[0004] While research on proteins for atopic dermatitis skin repair has made some progress, several challenges remain. First, the complex isolation and purification processes and high production costs of natural proteins limit their large-scale application. Second, while unmodified natural proteins can improve skin function, their effectiveness is still very limited, making it difficult to fully meet the clinical needs of patients with skin diseases such as atopic dermatitis. Therefore, it is necessary to find better ways to overcome these limitations. Summary of the Invention
[0005] The present invention aims to provide a novel recombinant protein, FCN1A-QTY-COIA1, for the treatment of atopic dermatitis. The existing FCN1A_XENLA and COIA1_HUMAN proteins are modified and redesigned through synthetic biology, retaining their original functional regions to exert their biological functions. Appropriate mutations are then made to some of the non-coding regions at their initial ends. The modified FCN1A_XENLA and COIA1_HUMAN proteins are then linked using a flexible protein linker (GGGGSGGGGSGGGGS), resulting in a novel recombinant protein, FCN1A-QTY-COIA1, with the ability to repair atopic dermatitis lesions. This invention utilizes synthetic biology to obtain a target protein with atopic dermatitis treatment function on a large scale, and can also reduce the production cost of natural protein-based dermatitis repair pharmaceutical preparations.
[0006] The above technical objectives of the present invention are achieved through the following technical solutions:
[0007] A novel recombinant protein FCN1A-QTY-COIA1, characterized by comprising partial amino acid sequences of FCN1A_XENLA and COIA1_HUMAN proteins, wherein leucine at positions 9, 10, 11, 37, 40, 79, 91, 113, 131, and 133 of the FCN1A_XENLA protein are all mutated to hydrophilic glutamines, for a total of 10 hydrophobic amino acid sites; and leucine at positions 7, 9, 13, 19, 26, 30, 48, 49, 57, and 63 of the COIA1_HUMAN protein are all mutated to glutamines, for a total of 10 hydrophobic amino acid sites. The amino acid sequence of the novel recombinant protein is shown in SEQ ID NO.1.
[0008] A recombinant expression vector, characterized by comprising the above-mentioned novel recombinant protein FCN1A-QTY-COIA1.
[0009] Preferably, the vector is selected from a plasmid vector, a phage vector or an animal or plant virus vector.
[0010] A recombinant bacterium, characterized by expressing the novel recombinant protein FCN1A-QTY-COIA1.
[0011] Preferably, the recombinant bacteria is selected from one of Escherichia coli, Bacillus subtilis, Pichia pastoris or Saccharomyces cerevisiae.
[0012] The use of the above-mentioned novel recombinant protein FCN1A-QTY-COIA1 in the preparation of pharmaceutical preparations for the treatment of atopic dermatitis.
[0013] Preferably, the pharmaceutical preparation further comprises a pharmaceutically acceptable carrier or adjuvant.
[0014] Preferably, the pharmaceutical preparation is a medicine for external use.
[0015] Preferably, the dosage form of the external medicine is a solution, cream or gel.
[0016] The application of the novel recombinant protein FCN1A-QTY-COIA1 in the preparation of in vitro skin fibroblast proliferation or migration promoting reagents, including experimental biochemical reagents.
[0017] In summary, the present invention has the following beneficial effects:
[0018] 1. While retaining the existing biological functions of the FCN1A_XENLA and COIA1_HUMAN proteins, the present invention modifies the amino acid sequences of some non-functional regions to enhance hydrophilicity through hydrophilic mutation and fusion via flexible linkers. This results in a novel recombinant protein with enhanced hydrophilicity, named FCN1A-QTY-COIA1. The specific amino acid sequence of the novel recombinant protein is shown in SEQ ID NO. 1. This novel recombinant protein has significant atopic dermatitis repair effects and enhanced hydrophilicity.
[0019] 2. In early experiments, the novel recombinant protein was tested for biocompatibility and atopic dermatitis repair properties. The results showed that the novel recombinant protein FCN1A-QTY-COIA1 has the ability to repair atopic dermatitis and has promising value for basic research and clinical applications. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following is a brief introduction to the drawings required for the embodiments or the description of the prior art. In all drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the various elements or parts are not necessarily drawn according to the actual scale. Obviously, the drawings described below are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can also be obtained based on these drawings without inventive work.
[0021] Figure 1 This is the electrophoretic band diagram of the novel recombinant protein FCN1A-QTY-COIA1 prepared by the present invention;
[0022] Figure 2 Electrophoretic band patterns of recombinant proteins with mutations at other different sites designed in the present invention;
[0023] Figure 3The advanced structure prediction results of the novel recombinant protein FCN1A-QTY-COIA1 designed for the present invention;
[0024] Figure 4 The hydrophilicity prediction results of the novel recombinant protein FCN1A-QTY-COIA1 designed for the present invention;
[0025] Figure 5 HEK293T cell toxicity assay of the novel recombinant protein FCN1A-QTY-COIA1 designed for the present invention;
[0026] Figure 6 L929 cell cytotoxicity assay of the novel recombinant protein FCN1A-QTY-COIA1 designed for the present invention;
[0027] Figure 7 This is an animal experiment on atopic dermatitis repair using the novel recombinant protein FCN1A-QTY-COIA1 designed for the present invention. DETAILED DESCRIPTION
[0028] To make the purpose, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.
[0029] As used herein, "and / or" includes any and all combinations of one or more of the associated listed items.
[0030] Herein, "plurality" means two or more than two, ie, it includes two, three, four, five, etc.
[0031] It should be noted that, in this document, the terms "comprises," "includes," or any other variations thereof are intended to encompass non-exclusive inclusion, such that a process, method, article, or apparatus comprising a series of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus. In the absence of further limitations, an element defined by the phrase "comprising a ..." does not exclude the presence of other identical elements in the process, method, article, or apparatus comprising the element.
[0032] As used in this specification, the term "about" typically means + / - 5% of the stated value, more typically + / - 4% of the stated value, more typically + / - 3% of the stated value, more typically + / - 2% of the stated value, even more typically + / - 1% of the stated value, and even more typically + / - 0.5% of the stated value.
[0033] In this specification, some embodiments may be disclosed in a format of being within a certain range. It should be understood that such description of "being within a certain range" is only for convenience and brevity and should not be interpreted as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered to have specifically disclosed all possible sub-ranges and independent numerical values within this range. For example, the range The description of should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within this range, for example, 1, 2, 3, 4, 5, and 6. The above rules apply regardless of the breadth of the range.
[0034] Materials and reagents:
[0035] HEK293T cells were purchased from the Cell Bank of the Type Culture Collection of the Chinese Academy of Sciences, catalog number: SCSP-502, and the cell name in Chinese is "human embryonic kidney cells." L929 cells were purchased from the Cell Bank of the Type Culture Collection of the Chinese Academy of Sciences, catalog number: SCSP-5039, and the cell name in Chinese is "mouse fibroblasts." Six-week-old male Balb / c mice were purchased from the Experimental Animal Center of Nantong University.
[0036] Escherichia coli BL21 (DE3) competent cells were purchased from Beijing Solebow Technology Co., Ltd.
[0037] The pET-22b(+) plasmid is a commercially available E. coli expression vector purchased from Beijing Qingke Biotechnology Co., Ltd. The vector tags are N-pelB and C-His, and the vector resistance is ampicillin.
[0038] Restriction endonucleases NdeI and XhoI were purchased from NEB (Beijing) Co., Ltd. His-tag protein purification resin (nickel column) was purchased from Shanghai Lianmai Bioengineering Co., Ltd., catalog number LM-616. IPTG, ampicillin, and DMSO (dimethyl sulfoxide) were all purchased from Beijing Solebold Technology Co., Ltd. DMEM medium was purchased from GIBCO. CCK8 reagent was purchased from Chongqing Baoguang Technology Co., Ltd. and used according to the reagent instructions.
[0039] Culture medium:
[0040] Each liter of LB medium contains: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, and the pH is adjusted to 7.0.
[0041] Preparation: Dissolve 5g yeast extract, 10g tryptone, and 10g sodium chloride in 950mL of double-distilled water. Adjust the pH to 7.0 with sodium hydroxide solution and dilute to 1L with double-distilled water. If preparing a solid medium, add agar at 1.5g / 100mL. Autoclave at 121°C for 30 minutes.
[0042] Experimental reagents not otherwise specified in the present invention are all conventional reagents in the art and can be prepared according to conventional methods in the art or purchased from relevant reagent suppliers; experimental methods not otherwise specified are all conventional methods in the art and reference can be made to relevant experimental manuals, such as molecular cloning experimental manuals or instructions from relevant reagent manufacturers.
[0043] Example 1
[0044] 1. FCN1A-QTY-COIA1 gene design and protein expression
[0045] A gene was designed, the nucleotide sequence of which is shown in SEQ ID NO.2, with a total length of 1374 bp. This gene encodes a protein, the amino acid sequence of which is shown in SEQ ID NO.1, with a total length of 458 amino acids, and is designated as the recombinant protein FCN1A-QTY-COIA1. During synthesis of this gene, an NdeI restriction site (CATATG) and an XhoI restriction site (CTCGAG) were added to the 5' and 3' ends of the gene, respectively. The gene sequence with the restriction sites is shown in SEQ ID NO.3. The synthesized gene was sequenced and verified, and the gene with the correct sequence was used for subsequent vector construction. The specific sequences involved are shown in Table 1.
[0046] Table 1
[0047]
[0048]
[0049]
[0050] 2. Vector Construction
[0051] In this embodiment, the pET-22b(+) plasmid is preferably used as an expression vector. The pET-22b(+) plasmid and the target gene (SEQ ID NO. 1) are double-digested with restriction endonucleases NdeI and XhoI, respectively, and the target gene is then ligated into the pET-22b(+) vector via ligation. It is understood that this embodiment can also be implemented using phage vectors or animal and plant virus vectors.
[0052] 3. Recombinant Bacterial Transformation
[0053] (1) Take out the competent Escherichia coli BL21 (DE3) cells (Beijing Solebio) from the -80℃ freezer and place them on ice for 5 minutes.
[0054] (2) After the glycerol containing the BL21(DE3) competent cells has melted, add the competent cells to the ligation product, pipette and release the solution three times to mix thoroughly, and place on ice for 30 minutes.
[0055] (3) Quickly wipe the tube wall dry with absorbent paper, then heat shock at 42°C for 90 seconds, and immediately place on ice for 2 minutes.
[0056] (4) Add 800 μL of LB liquid culture medium under sterile conditions and culture at 37°C, 150 rpm for 45 min.
[0057] (5) Collect the cells by centrifugation at 8000 rpm for 5 min, discard part of the supernatant, and use the remaining 100 μL of supernatant to resuspend the E. coli. Then, spread it evenly on LB solid culture medium containing 100 μg / mL ampicillin, place it in a 37°C incubator, and invert and culture for about 12 h.
[0058] (6) A single clone was picked and inoculated into liquid LB medium containing 100 μg / mL ampicillin. After incubation at 37°C for about 13 h, positive clones were identified.
[0059] This embodiment preferably uses Escherichia coli as the recombinant bacteria. It is understood that this embodiment can also be implemented using Bacillus subtilis, P. cerevisiae, or Saccharomyces cerevisiae strains.
[0060] 4. Protein Expression and Purification
[0061] (1) Inoculation: Prepare liquid LB medium and sterilize it. Place the sterilized liquid LB medium in a clean bench and cool it to room temperature. Add ampicillin to the LB medium in the clean bench and mix it to a final concentration of 100 μg / mL. Then, inoculate Escherichia coli (positive clone) containing the target gene plasmid into the LB medium at a volume of 200 μL / L. Then, place the LB medium in a shaker and culture it at a speed of 170 rpm and a temperature of 37°C for 8 h.
[0062] (2) Induction: After 8 h of shaking culture, 2 mL of bacterial solution was taken out and its OD was measured by spectrophotometer. 600 When the OD value of the bacterial solution 600When the value reaches 0.6-0.8, IPTG is added to the bacterial solution to a final concentration of 200 μL / mL, and then the bacterial solution is cultured at 37°C and 170 rpm for 8 hours.
[0063] (3) Purification: After adding IPTG and culturing for 8 hours, the bacterial solution is centrifuged at 4°C and 8000 rpm for 5 minutes. After centrifugation, the supernatant is removed, and the precipitate (i.e., E. coli) is retained.
[0064] (4) Ultrasonic disruption: After ultrasonic disruption of the E. coli, centrifugation is performed. The precipitate obtained by centrifugation contains the target protein. The precipitate obtained by centrifugation is first washed with washing solution I (50 mmol / L Tris-HCl, 1 mol / L urea, 10 mL / L Triton X-100), and then washed with washing solution II (50 mmol / L Tris-HCl, 2 mol / L urea, 5 mL / L Triton X-100). Finally, the inclusion body solution (50 mmol / L Tris-HCl, 8 mol / L urea, 100 mmol / L NaCl) is used for dissolution.
[0065] (5) Finally, the above inclusion body solution is subjected to gradient renaturation as follows: the inclusion body solution is placed in a dialysis bag (Beijing Solaybao Technology Co., Ltd., item number: YA1071), and then the dialysis bag is sequentially placed in 6M, 4M, 2M, 1M, and 0.5M urea solutions for gradient renaturation, with 3 hours of renaturation at each urea concentration. The renaturation is performed at 4°C. After renaturation, the solution is purified by His-tag protein purification resin (nickel column, Shanghai Linkmate) to obtain the recombinant protein FCN1A-QTY-COIA1 (the gene sequence of the target protein is designed to add a histidine tag (HHHHHHHHHH)). After purification, polyacrylamide gel electrophoresis (SDS-PAGE) is performed to identify whether the purified target protein is successfully obtained. The results are shown in Figure 1 , and the target protein with a size of about 49.6 kDa is obtained.
[0066] The detailed information of the novel recombinant protein FCN1A-QTY-COIA1 is as follows:
[0067]
[0068] The high-level structure of the recombinant protein FCN1A-QTY-COIA1 prepared in this example is predicted, and the results are shown in Figure 3 . The high-level structure of the novel recombinant protein FCN1A-QTY-COIA1 contains a large number of "α-helix" secondary structures.
[0069] Furthermore, the hydrophilicity prediction of the novel recombinant protein FCN1A-QTY-COIA1 prepared in this example was performed, and the results are shown in FIG. Figure 4 . Figure 4 The horizontal axis represents the protein's amino acid sequence; the vertical axis represents hydrophilicity, with larger values indicating greater hydrophobicity and negative values indicating hydrophilicity. The total hydrophilicity value of the novel recombinant protein FCN1A-QTY-COIA1 (-0.689) is lower than that of the native proteins FCN1A_XENLA (-0.505) and COIA1_HUMAN (0.244), indicating that the novel recombinant protein with atopic dermatitis repair function prepared in this invention is more hydrophilic.
[0070] In the preliminary experiments of the present invention, recombinant proteins with 16, 18, 22, and 24 site mutations were constructed, and the construction, expression, and purification steps were consistent with those of the novel recombinant protein FCN1A-QTY-COIA1. Figure 2 The results showed that the recombinant proteins with mutations at amino acid sites 16, 18, 22, and 24 could not be effectively expressed, and no obvious protein expression was observed on the SDS-PAGE electrophoresis bands.
[0071] Example 2
[0072] Advanced structure prediction of the novel recombinant protein FCN1A-QTY-COIA1
[0073] The entire amino acid sequence of the novel recombinant protein FCN1A-QTY-COIA1 was input into the Alphafold3 online prediction website (https: / / alphafoldserver.com / ), submitted, and prediction began.
[0074] Open the prediction result file using Pymol software, such as Figure 3 As shown, the obtained high-level structure prediction results are analyzed.
[0075] Example 3
[0076] Analysis of the overall hydrophilicity value of the new recombinant protein FCN1A-QTY-COIA1
[0077] Enter the entire amino acid sequence of the new recombinant protein FCN1A-QTY-COIA1 into the online website https: / / web.expasy.org / protscale / , click Submit and Prediction, and you will get the hydrophilicity analysis results of different amino acid sites of the new recombinant protein.
[0078] like Figure 4As shown in the hydrophilicity results graph, the horizontal axis represents amino acid sites, and the vertical axis represents hydrophilicity. Positive vertical values indicate hydrophobicity, while negative vertical values indicate hydrophilicity. Larger vertical values indicate more hydrophobicity, while smaller vertical values indicate more hydrophilicity. The more hydrophilic the protein, the better the moisturizing function of its finished formulation.
[0079] Example 4
[0080] Cytotoxicity evaluation of the novel recombinant protein FCN1A-QTY-COIA1
[0081] The recombinant protein FCN1A-QTY-COIA1 prepared in Example 1 was tested on normal HEK293T and L929 cells by CCK8 assay to evaluate its biosafety.
[0082] Method steps:
[0083] (1) In a 96-well plate, 1×10 4 Human embryonic kidney HEK293T cells were cultured in DMEM medium, and five experimental groups and one control group were set up, with three replicate wells in each group.
[0084] (2) After culturing the cells at 37°C for 24 hours, the novel recombinant protein FCN1A-QTY-COIA1 was added to the wells of the experimental groups (experimental wells) so that the final concentrations of the novel recombinant protein FCN1A-QTY-COIA1 in the five experimental groups were 10 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL, and 400 μg / mL, respectively. No novel recombinant protein RB1-PDL1 was added to the cells in the wells of the control group (control wells).
[0085] (3) Continue to culture at 37℃ for 24h, and use CCK8 method to detect the number of living cells. The steps are as follows: add 10μL CCK8 reagent to each well and incubate at 37℃ for 3h. Then use a microplate reader to measure the absorbance of each well at 450nm.
[0086] See the results Figure 5 , Figure 5 It shows that when the concentration of the new recombinant protein FCN1A-QTY-COIA1 reaches 400μg / mL, the survival rate of HEK293T cells still remains above 90%, indicating that the new recombinant protein FCN1A-QTY-COIA1 has no obvious cytotoxicity and no obvious inhibitory or killing effect on normal cells, and can be further applied to in vitro and in vivo research, diagnosis and treatment. At the same time, Figure 5 The experimental results also showed that compared with the unmodified FCN1A_XENLA and COIA1_HUMAN proteins, the new recombinant protein FCN1A-QTY-COIA1 was significantly less cytotoxic to HEK293T cells.
[0087] The cytotoxicity assay procedure for L929 cells is consistent with that for HEK293T cells.
[0088] like Figure 6 The results of the cytotoxicity experiment of L929 cells also showed that compared with the unmodified FCN1A_XENLA and COIA1_HUMAN proteins, the new recombinant protein FCN1A-QTY-COIA1 has less cytotoxicity and better biocompatibility for normal cells L929.
[0089] Example 5
[0090] Atopic Dermatitis Treatment Trial
[0091] Atopic dermatitis mouse model: Female mice were randomly divided into three groups (n=3). 1.5% DNCB (2,4-dinitrochlorobenzene) and 1% DNCB were applied to both ears of the mice for three consecutive days, then every other day. The model was established after 10 days.
[0092] Skin repair experiment: One of the three groups of mice was randomly selected to receive a 3% recombinant protein FCN1A-QTY-COIA1 solution applied to both ears. Another group received a glucocorticoid (dexamethasone ointment). A third group served as a control group and received no treatment. Treatment was continued for 14 consecutive days (applying the solution once daily at a scheduled time). Photos were taken before each application to observe the effects of ear atopic dermatitis repair.
[0093] The recombinant protein's ability to treat atopic dermatitis was evaluated using a mouse bilateral ear atopic dermatitis model. The recombinant protein solution was applied to the dermatitis site. The ears of the control mice did not recover after two weeks. However, the ears of the mice treated with 3% recombinant protein FCN1A-QTY-COIA1 solution essentially recovered, with bilateral recovery significantly better than in the control and glucocorticoid groups ( Figure 7 The results showed that the recombinant protein FCN1A-QTY-COIA1 can effectively improve skin damage caused by atopic dermatitis.
[0094] The embodiments of the present invention are described above in conjunction with the accompanying drawings, but the present invention is not limited to the above-mentioned specific implementation methods. The above-mentioned specific implementation methods are merely illustrative and not restrictive. Under the guidance of the present invention, ordinary technicians in this field can also make many forms without departing from the scope of protection of the present invention and the claims, all of which are protected by the present invention.
Claims
1. A novel recombinant protein FCN1A-QTY-COIA1, characterized in that: The novel recombinant protein comprises partial amino acid sequences of FCN1A_XENLA and COIA1_HUMAN proteins, wherein the leucine residues at positions 9, 10, 11, 37, 40, 79, 91, 113, 131, and 133 of the FCN1A_XENLA protein are all mutated to hydrophilic glutamines, for a total of 10 hydrophobic amino acid sites; and the leucine residues at positions 7, 9, 13, 19, 26, 30, 48, 49, 57, and 63 of the COIA1_HUMAN protein are all mutated to glutamines, for a total of 10 hydrophobic amino acid sites. The amino acid sequence of the novel recombinant protein is shown in SEQ ID NO.
1.
2. A recombinant expression vector, characterized in that: It comprises the novel recombinant protein FCN1A-QTY-COIA1 according to claim 1.
3. The recombinant expression vector according to claim 2, wherein The vector is selected from a plasmid vector, a phage vector or an animal or plant virus vector.
4. A recombinant bacterium, characterized in that Expressing the novel recombinant protein FCN1A-QTY-COIA1 according to claim 1.
5. The recombinant bacterium according to claim 4, wherein The recombinant bacteria is selected from one of Escherichia coli, Bacillus subtilis, Pichia pastoris or Saccharomyces cerevisiae.
6. Use of the novel recombinant protein FCN1A-QTY-COIA1 according to claim 1 in the preparation of pharmaceutical preparations for treating atopic dermatitis.
7. The use according to claim 6, characterized in that The pharmaceutical preparation further includes a pharmaceutically acceptable carrier or adjuvant.
8. The use according to claim 6, characterized in that The pharmaceutical preparation is a medicine for external use.
9. The use according to claim 8, characterized in that The dosage form of the external medicine is solution, cream or gel.
10. Use of the novel recombinant protein FCN1A-QTY-COIA1 according to claim 1 in the preparation of an in vitro reagent for promoting proliferation or migration of skin fibroblasts, including experimental biochemical reagents.