CN108586616A - A kind of fusion protein and its preparation method and application promoting endothelium reparation - Google Patents

A kind of fusion protein and its preparation method and application promoting endothelium reparation Download PDF

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CN108586616A
CN108586616A CN201810135751.2A CN201810135751A CN108586616A CN 108586616 A CN108586616 A CN 108586616A CN 201810135751 A CN201810135751 A CN 201810135751A CN 108586616 A CN108586616 A CN 108586616A
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fusion protein
seq
cadherin
mfp
amino acid
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CN108586616B (en
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黄俊丽
杨东川
王贵学
邱菊辉
赵银瓶
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Chongqing University
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Abstract

The invention belongs to gene engineering technology fields, and in particular to a kind of fusion protein and its preparation method and application promoting endothelium reparation.The present invention promotes the amino acid sequence of the fusion protein of endothelium reparation as shown in SEQ ID NO.1.The fusion protein provided by the invention for promoting endothelium reparation, endothelial cell and endothelial progenitor cells can efficiently and be specifically promoted to adhere to, it can promote the proliferation of endothelial cell, and adherency and the endothelial inflammation of smooth muscle cell will not be increased, close connection and the NO releases for being conducive to endothelial cell, to accelerate the endothelialization of blood vessel.The fusion protein or its recombinant vector prepared can be applied in preparing the biomaterial for promoting endothelium reparation, to clinically there is important meaning.

Description

A kind of fusion protein and its preparation method and application promoting endothelium reparation
Technical field
The invention belongs to gene engineering technology fields, and in particular to it is a kind of promote endothelium reparation fusion protein and its preparation Methods and applications.
Background technology
Cardiovascular implant material is closely related with quality of life with human disease treatment.Current clinically used angiocarpy It includes artificial blood vessel and intravascular stent to be implanted into material mainly.Artificial blood vessel easily occurs thrombus, increases due to inorganizable power of regeneration The problems such as raw and calcification；Vascular implantation material based on endovascular stent, is in direct contact with blood and tube wall tissue in vivo, And it is the main problem in clinic to cause mechanical damage, thrombosis and the restenosis occurred thereafter to implant site.These are asked Topic is mostly derived from the surface characteristic of implantation material.Therefore clinical application proposes the surface characteristic for being implanted into biomaterial higher It is required that.How material surface is entered to cardiovascular plant and carry out multifunctional modification, so that rack surface is had and promote endothelialization, antithrombotic shape At, functions such as anti-inflammatory, anti-endometrial hyperplasia, it is of great significance to the exploitation of cardiovascular implantation material with applying.Currently, painstaking effort Pipe implantation material surface is modified generally use plasma spray coating and layer-by-layer.However these technologies are with larger Limitation, as plasma spray technology is to the size of sample, shape need is stringent and operating process is complicated；LBL self-assembly is poly- Electrolytic multi-layer film deposits the material surface that need to be charged, and the structural stability formed is bad, and carrying out surface with this kind of technology changes The long-term patency rate of the intravascular stent of property is still to be improved.
Endovascular stent implantation is the interventional therapeutic technique to grow up on the basis of percutaneous transluminal angiography, In wide clinical application, the only 1 year case load for receiving endovascular stent implantation in U.S. l0 ten thousand nearby.Recent study Emphasis is endothelium reparation, inhibition thrombosis and the smooth muscle cell proliferation for promoting stenter to implant section blood vessel, is inhibited to reach The purpose of endometrial hyperplasia.
Intravascular tissue engineering implantation material is without specificity at present, in the same of the endometrial hyperplasia for inhibiting smooth muscle abnormal in other words When can also inhibit the biological behaviour and function of endothelial cell mostly, postpone endothelialization.
Therefore, there is an urgent need to develop new implantation material surface modification technology, cardiovascular implantation material is further promoted Clinical efficacy and application quality.
Invention content
In view of this, the purpose of the present invention is to provide a kind of fusion protein promoting endothelium reparation, the fusion protein energy Promote efficiently and specifically endothelial cell adherency, the proliferation of endothelial cell can be promoted, be conducive to the close connection of endothelial cell It is discharged with NO.
To achieve the above object, the technical scheme is that：
Promote the fusion protein of endothelium reparation, which is characterized in that the amino acid sequence of the fusion protein such as SEQ ID Shown in NO.1.
Further, the fusion protein is merged by the extracellular domain 1,2 (EC1-2) and Mfp-5 of VE-cadherin It arrives.
Intravascular tissue engineering implantation material is without specificity at present, in the same of the endometrial hyperplasia for inhibiting smooth muscle abnormal in other words When can also inhibit the biological behaviour and function of endothelial cell mostly, postpone endothelialization.But since VE-cadherin is only to deposit It is a kind of adhesion molecule of endothelial cell, so the fusion protein that the present invention is built has the promotion inner skin cell viscosity of specificity The effects that attached, to accelerate the endothelialization of blood vessel.
Further, the amino acid sequence such as SEQ ID NO.2 of the extracellular domain 1,2 (EC1-2) of the VE-cadherin It is shown；The amino acid sequence of the Mfp-5 is as shown in SEQ ID NO.3.
Mfp-5 is to be currently known the strongest mussel byssus protein of adhesive capacity.Mussel byssus protein includes a variety of different Byssus protein type, at least 8 kinds, but containing there are many be rich in L-3,4- dihydroxy-phenylalanine (3,4- dihydroxyphenylalanine,DOPA).Wherein Mfp-3 and Mfp-5 is two kinds of highest protein of DOPA contents in Mfp, It is predominantly located at the interface of focal adhension and solid substrate surface, and the mfp-5 adhesiving effects that the present invention selects are good, yield is high, and And the problems such as not will produce cytotoxicity, immunological rejection.Mfp-5 can be adhered strongly to the arbitrary surface of solids in wet environment, In pH2.6 environment, the adhesion energy between Mfp-5 and mica surface can reach -13.7mJ/m2, between Mfp-5 and Mfp-5 Cohesive energy can reach -2.5mJ/m2。
Adhesion molecules of the VE-cadherin as endothelial-cell specific is by VE-cadherin extracellular domains 1 Interaction forms trans- dimer and Adjacent endothelial cells is closely connected, and is maintaining the integrity of endothelium and new vessels Occur and plays particularly important effect in generating process.
The holder energy efficient capture endothelial cell and endothelial progenitor cells of VE-cadherin antibody coatings, 3d after stenter to implant Good endothelialization is shown, more preferable (bibliography Comparison of more with obvious effects than CD34 antibody scaffold endothelialization and neointimal formation with stents coated with antibodies against CD34and vascular endothelial-cadherin).Therefore the present invention selects VE- Cadherin EC1-2 and Mfp-5 construction of fusion protein has reached the effect for promoting vessel endothelialisation.
The fusion protein of the present invention has good adhesion property to solid substrate, is easy coating (load), can specificity Ground promotes the adherency of endothelial cell and endothelial progenitor cells, and will not increase adherency and the endothelial inflammation of smooth muscle cell.
The second object of the present invention is to provide a kind of preparation method of above-mentioned fusion protein, include the following steps：
1) gene order of purpose fusion protein is synthesized：In the nucleotide sequence SEQ ID of VE-cadherin EC1-2 The junctions nucleotide sequence SEQ ID NO.5 of NO.4 and Mfp-5 add linker, then by recombinase by two target sequences Fusion, obtains the sequence as shown in SEQ ID NO.7；
2) construction recombination plasmid：The gene order that step 1) obtains is cloned by ligase in carrier, load must be recombinated Body, and prepare transformant and extraction recombinant plasmid；
3) expression of recombinant plasmid：Recombinant plasmid transformed to the host strain that step 2) obtains is obtained into recombinant bacterial strain, then through luring Expression is led, fusion protein is obtained.
Further, the nucleotide sequence of the step 1) linker is as shown in SEQ ID NO.6.
In addition to prokaryotic expression, after replacing different expression vectors, other a variety of corresponding expression systems can also express this Fusion protein, including yeast, insect cell, mammalian cell.After expressing fusion protein, dye method can be examined with SDS-PAGE and is exempted from Epidemic disease blotting is identified, the methods of BCA methods or enzyme-linked immunosorbent assay (ELISA) can be used to measure the albumen concentration of purifying.
Demonstrate fusion protein using Spr instrument and atomic force microscope (AFM) has good adhesion work to a variety of substrates Energy.The adhesion experiment and antibody neutralization test of endothelial cell and endothelial progenitor cells prove that the fusion protein that the present invention is built can be efficiently And specifically promote endothelial cell adherency, moreover, fusion protein can promote the proliferation of endothelial cell, and be conducive to endothelial cell It is close connection and NO release etc. functions.
A kind of specific preparation method of fusion protein, includes the following steps：
1) gene order of purpose fusion protein is synthesized：Obtain the DNA sequences of coding VE-cadherin EC1-2 and Mfp-5 Row, the acquisition of DNA sequence dna are added by conventional RNA extractions, reversion and PCR synthetic technologys in the junctions the two DNA respectively Two target DNA fragments are fused to one section by 15 homologous bases as linker, then by In-fusion recombinases, obtain as Sequence shown in SEQ ID NO.7；
2) construction recombination plasmid：The gene order that step 1) obtains is cloned by T4 ligases in carrier PET32a, Recombinant vector is obtained, and prepares transformant and extraction recombinant plasmid；
3) expression and purification of recombinant plasmid：It is thin that the recombinant plasmid that step 2) is obtained imports Escherichia coli DE3 (BL21) In born of the same parents, then the extraction purification fusion protein after induction makes expressing fusion protein.
The preparation method of the present invention is simple, and wherein carrier PET32a includes the promoter for driving gene expression, egg The breeding of white matter translation initiation signal and antibiotic resistance genes in favor of plasmid in bacterium, and carry histidine tag Conducive to the purifying of following protein.
The third object of the present invention is to provide a kind of above-mentioned fusion protein and is preparing the biomaterial for promoting endothelium reparation In application.
Further, the biomaterial includes rack surface coating, artificial blood vessel's surface modification, protein active medicine Object.
The present invention also aims to provide the recombinant vector obtained in a kind of above-mentioned preparation method.
The present invention also aims to provide a kind of recombinant vector to prepare blood vessel endothelium injury repair medicine or examination Application in agent.The biomaterial includes rack surface coating, artificial blood vessel's surface modification, protein active drug.
The present invention also aims to provide the extracellular domain 1,2 (EC1-2) of VE-cadherin a kind of and Mfp-5 to exist Prepare the application in blood vessel endothelium injury repair medicine or reagent.
Further, the amino acid sequence such as SEQ ID NO.2 of the extracellular domain 1,2 (EC1-2) of the VE-cadherin It is shown；The amino acid sequence of the Mfp-5 is as shown in SEQ ID NO.3.
The beneficial effects of the present invention are：
1) fusion protein provided by the invention for promoting endothelium reparation, can efficiently and specifically promote endothelial cell and interior Skin progenitor cells adhere to, and can promote the proliferation of endothelial cell, and will not increase adherency and the endothelial inflammation of smooth muscle cell, be conducive to The close connection and NO releases of endothelial cell, to accelerate the endothelialization of blood vessel.
2) preparation method of the invention is simple, and wherein carrier PET32a includes the promoter for driving gene expression, The breeding of protein translation initial signal and antibiotic resistance genes in favor of plasmid in bacterium, and carry histidine mark Label are conducive to the purifying of following protein.
3) fusion protein of the invention or its recombinant vector prepared, which can be applied, is preparing the biological material for promoting endothelium reparation In material, to clinically there is important meaning.
Description of the drawings
Fig. 1 is fusion protein expression plasmid.
Fig. 2 is the purifying situation of fusion protein.MW is marker, and M is that the mussel protein Mfp-5, VE-M of recombination are fusions Albumen, VE are the VE-cadherin EC1-2 of recombination.
Fig. 3 is fusion protein has good Adhering capacity in a variety of substrates, can form uniform coating.
Fig. 4 is the adherency situation on endothelial cell fusion protein coating.A is endothelial cell sticking machine schematic diagram, and B, C are For endothelial cell in the adherency number of different base, D is that endothelial cell sprawls situation in different base.
Fig. 5 is the adherency situation on endothelial progenitor cells fusion protein coating.A is the mirror of the endothelial progenitor cells of in-vitro separation Fixed, B, C are adherency number of the endothelial progenitor cells in different base.
Fig. 6 illustrates that fusion protein specifically promotes the adherency of endothelial cell by VE-cadherin EC1-2.A, B is smooth Myocyte adheres to situation.C is antibody neutralization test schematic diagram, and D, E are antibody neutralization test result.
Fig. 7 is the influence of the endogenous expression VE-cadherin of fusion protein Human Umbilical Vein Endothelial Cells.A is immunofluorescence, B qPCR As a result, C is WB as a result, fusion protein can promote the endogenous expression VE-cadherin of endothelial cell to increase.
Fig. 8 is the proliferative conditions of endothelial cell on different substrates.
Fig. 9 A are the influence that different base Human Umbilical Vein Endothelial Cells supernatant expresses eNOS levels, and B, C are THP-1 pairs in different base The adherency situation of endothelial cell.
Figure 10 is the close-connected influence of fusion protein Human Umbilical Vein Endothelial Cells.A is the immunofluorescence of F-actin, and B, C are not With the penetrating implementations of monolayer endothelial cell in substrate, D, E, F be respectively Tight junction protein occludin, Claudin-5 and The transcriptional level situation of ZO-1.
Specific implementation mode
Hereinafter reference will be made to the drawings, and the preferred embodiment of the present invention is described in detail.Tool is not specified in preferred embodiment The experimental method of concrete conditions in the establishment of a specific crime, usually according to normal condition, illustrated embodiment are to preferably be said to present disclosure It is bright, but be not that present disclosure is only limitted to illustrated embodiment.So those skilled in the art are according to foregoing invention Content carries out nonessential modifications and adaptations to embodiment, still falls within protection scope of the present invention.
The structure of 1 fusion protein expression plasmid of embodiment
The present invention extracts total serum IgE using Trizol (Takara companies) from Human umbilical vein endothelial cells (HUVECs), then RNA is inverted to cDNA with Reverse Transcriptase kit.Amplification is reacted by PCR with specific primer again and obtains required VE- Cadherin (EC1-2) segment.Mfp-5 obtains required Mfp-5 segments since source is not easy, by chemically synthesized method. 15 homologous bases are respectively added in VE-cadherin (EC1-2) and Mfp-5 sequences junction, to use In-fusion recombinases will VE-cadherin (EC1-2) and Mfp-5 sequences are merged, and are then cloned into carrier PET32a, are seen by T4 ligases Fig. 1 fusion protein expression plasmid schematic diagrames.
The expression and purification of 2 fusion protein of embodiment
After the structure for completing expression plasmid, plasmid is imported in Escherichia coli DE3 (BL21) cell, according to required egg White amount, the appropriate Bacillus coli cells comprising destination gene expression carrier of culture to OD600 are 0.6-0.8, and 0.05mM is added IPTG induce 5-8h to make expressing fusion protein at 28 DEG C, 8000rpm centrifugations 20min collects thalline at 4 DEG C, then uses ultrasound Broken instrument ice-bath ultrasonic is crushed Bacillus coli cells, and supernatant is collected by centrifugation, is carried by His labels using nickel column after 0.45um filterings Purified fusion albumen is taken, obtained albumen ice bath is dialysed to be preserved into 1%HAc.The concentration of fusion protein is measured with BCA methods, and It is identified by immunoblotting, the purifying situation of fusion protein is shown in Fig. 2.
The adhesion experiment of embodiment 3 fusion protein and substrate
It is stainless in 316L to determine fusion protein by being positioned to fusion protein in conjunction with FITC fluorescent moleculars by the present invention Steel, the adherency of tri- kinds of different bases of PLCL and TC-PS and coating conditions put the addition of fusion protein solution in this experiment Have in 24 orifice plates of different base, incubation at room temperature for 24 hours, is taken pictures after ultrasonic cleaning with fluorescence microscope, and utilizes atomic force microscopy Mirror and water contact angle instrument characterize fusion protein coating.Fusion protein has been observed in real time in gold plaque simultaneously also by SPR instrument On adherency situation.
Three kinds of substrates have apparent uniform fluoresent coating after immersion, illustrate the fusion protein of present invention structure expression to solid There is body substrate good adhesive capacity, SPR instrument can also detect the binding signal of fusion protein and gold plaque substrate, preceding 200s gold The fusion protein amount about 100ng/cm that piece combines2, when flushing only has minute quantity to be rinsed in conjunction with untight albumen.Merge egg The water contact angle of white coating is changed compared to bare substrate, and hydrophily is more moderate, these the result shows that fusion protein at Work(adheres to substrate and forms uniform coating, sees Fig. 3.
4 fusion protein of embodiment specifically promotes endothelial cell to adhere to
The critical instance of the present invention is to demonstrate constructed fusion protein specifically endothelial cell can be promoted to adhere to.It is logical Endothelial cell and endothelial progenitor cells static state adhesion experiment are crossed, using the cells such as collagen and PLL typical binders as positive control, together Shi Liyong VE-cadherin and integrin antibody carries out specific neutralization test, and the thin of 30min observation each groups is incubated at 37 DEG C Born of the same parents adhere to number.
As a result see Fig. 4, Fig. 5, Fig. 6 and Fig. 7, adhesion experiment the results show that fusion protein constructed by the present invention compared to Collagen, PLL etc. can be obviously promoted the adherency of endothelial cell and endothelial progenitor cells, and the endothelial cell of fusion protein group adherency is about 4 times of negative control group are 1.5 times or more of collagen and PLL, and cell is sprawled also significantly more.And for endothelial progenitor cells, The cell number that fusion protein sticks is at least 2 times of other group.Neutralization test first passes through antibody in advance, and specificity closing is thin respectively Sites VE-cadherin and integrin on born of the same parents, it was demonstrated that fusion protein be by with the VE-cadherin on endothelial cell It specifically combines and promotes endothelial cell adherency, and immunofluorescence and the testing result of qPCR find endothelial cell endogenous The expression of VE-cadherin is significantly raised compared to other group.
5 fusion protein of embodiment is effectively facilitated endothelial cell proliferation
Another critical instance of the present invention is to demonstrate the fusion protein of structure to effectively facilitate the proliferation of endothelial cell.At this In one experiment, HUVECs is inoculated in 24 orifice plates for being placed with 316L stainless steel sheet materials, exposed sheet material is real as negative control The advance coating fusion protein of group is tested, is added after endothelial cell at 37 DEG C after culture for 24 hours, immunofluorescence dyeing Ki67 calculates Ki67 Positive endothelial cell percentage.
Proliferation experiment result show the Ki67 positive Endothelial Cells of fusion protein group obviously mostly with control group, and with notable Sex differernce is shown in Fig. 8.
6 fusion protein of embodiment reinforces the functions such as close connection and the NO releases of endothelial cell
The application of construction of fusion protein of the present invention is for promoting vessel endothelialisation, the function of fusion protein Human Umbilical Vein Endothelial Cells Influence it is particularly important, therefore the present invention has detected the shadow of the functions such as close connection and the NO releases of fusion protein Human Umbilical Vein Endothelial Cells It rings.As a result see Fig. 9 and Figure 10, in permeability experiment, advance coating fusion protein is in the cells Transwell, without any painting Layer is used as negative control, inoculates after HUVECs cultivate for 24 hours at 37 DEG C, the FITC-dextron of upper chamber addition 1mg/mL is 37 DEG C it is incubated 15min, the fluorescence intensity of lower room is detected with sepectrophotofluorometer, as a result, it has been found that the fluorescence intensity pole of fusion protein group Substantially less than other groups illustrate that the endothelial cell of fusion protein group connects the closest, the immunofluorescence dyeing result of skeleton It is consistent with this.Meanwhile the present invention has detected the transcription situation of 3 kinds of main tight junction proteins of endothelial cell, qPCR results Although the transcriptional level of display Occludin and Claudin-5 does not have significant change, the expression of ZO-1 obviously to raise.
Finally illustrate, the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although with reference to compared with Good embodiment describes the invention in detail, it will be understood by those of ordinary skill in the art that, it can be to the skill of the present invention Art scheme is modified or replaced equivalently, and without departing from the objective and range of technical solution of the present invention, should all be covered at this In the right of invention.
<110>University Of Chongqing
<120>A kind of fusion protein and its preparation method and application promoting endothelium reparation
<160> 7
<170> PatentIn version 3.3
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<213> Artificial
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<223>Fusion protein
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Thr Ala Val Asp Ala Asp Asp Pro Thr Val Gly Asp His Ala Ser
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Val Met Tyr Gln Ile Leu Lys Gly Lys Glu Tyr Phe Ala Ile Asp
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Asn Ser Gly Arg Ile Ile Thr Ile Thr Lys Ser Leu Asp Arg Glu
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Lys Gln Ala Arg Tyr Glu Ile Val Val Glu Ala Arg Asp Ala Gln
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Gly Leu Arg Gly Asp Ser Gly Thr Ala Thr Val Leu Val Thr Leu
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Tyr Tyr Pro Gly Asn Thr Tyr His Tyr His Ser Gly Gly Ser Tyr
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Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr Lys Asn Ser Gly Lys Tyr
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Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg Lys Gly Tyr Lys
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Lys Tyr Tyr Gly Gly Gly Ser Ser
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cgggtcgatg cagagacagg agacgtgttc gccattgaga ggctggaccg ggagaatatc 180
tcagagtacc acctcactgc tgtcattgtg gacaaggaca ctggcgaaaa cctggagact 240
ccttccagct tcaccatcaa agttcatgac gtgaacgaca actggcctgt gttcacgcat 300
cggttgttca atgcgtccgt gcctgagtcg tcggctgtgg ggacctcagt catctctgtg 360
acagcagtgg atgcagacga ccccactgtg ggagaccacg cctctgtcat gtaccaaatc 420
ctgaagggga aagagtattt tgccatcgat aattctggac gtattatcac aataacgaaa 480
agcttggacc gagagaagca ggccaggtat gagatcgtgg tggaagcgcg agatgcccag 540
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cgggtcgatg cagagacagg agacgtgttc gccattgaga ggctggaccg ggagaatatc 180
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ccttccagct tcaccatcaa agttcatgac gtgaacgaca actggcctgt gttcacgcat 300
cggttgttca atgcgtccgt gcctgagtcg tcggctgtgg ggacctcagt catctctgtg 360
acagcagtgg atgcagacga ccccactgtg ggagaccacg cctctgtcat gtaccaaatc 420
ctgaagggga aagagtattt tgccatcgat aattctggac gtattatcac aataacgaaa 480
agcttggacc gagagaagca ggccaggtat gagatcgtgg tggaagcgcg agatgcccag 540
ggcctccggg gggactcggg cacggccacc gtgctggtca ctctgcaaga cggtggtggt 600
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tcaggtggta gttatcacgg atccggctat catggaggat ataagggaaa gtattacgga 720
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gctagaaaat accatagaaa gggttacaag aagtattatg gaggtggtag cagttag 837

Claims (9)

1. promoting the fusion protein of endothelium reparation, which is characterized in that the amino acid sequence of the fusion protein such as SEQ ID NO.1 It is shown.
2. fusion protein according to claim 1, which is characterized in that the fusion protein is by the extracellular of VE-cadherin Structural domain 1,2 (EC1-2) and Mfp-5 merge to obtain.
3. fusion protein according to claim 2, which is characterized in that the extracellular domain 1,2 of the VE-cadherin (EC1-2) amino acid sequence is as shown in SEQ ID NO.2；The amino acid sequence of the Mfp-5 is as shown in SEQ ID NO.3.
4. the preparation method of claims 1 to 3 any one of them fusion protein, which is characterized in that include the following steps：
1) gene order of purpose fusion protein is synthesized：VE-cadherin EC1-2 nucleotide sequence SEQ ID NO.4 and The junctions nucleotide sequence SEQ ID NO.5 of Mfp-5 add linker, then are merged two target sequences by recombinase, Obtain the sequence as shown in SEQ ID NO.7；
2) construction recombination plasmid：The gene order that step 1) obtains is cloned by ligase in carrier, recombinant vector is obtained, and Prepare transformant and extraction recombinant plasmid；
3) expression of recombinant plasmid：Recombinant plasmid transformed to the host strain that step 2) obtains is obtained into recombinant bacterial strain, then through inducing table It reaches, obtains fusion protein.
5. preparation method according to claim 4, which is characterized in that the nucleotide sequence of the step 1) linker is such as Shown in SEQ ID NO.6.
6. application of claims 1 to 3 any one of them fusion protein in preparing the biomaterial for promoting endothelium reparation.
7. application according to claim 6, which is characterized in that the biomaterial includes rack surface coating, artificial blood Pipe surface trim, protein active drug.
The extracellular domain 1,2 (EC1-2) and Mfp-5 of 8.VE-cadherin is preparing blood vessel endothelium injury repair medicine or examination Application in agent.
9. application according to claim 8, which is characterized in that 1, the 2 (EC1- of extracellular domain of the VE-cadherin 2) amino acid sequence is as shown in SEQ ID NO.2；The amino acid sequence of the Mfp-5 is as shown in SEQ ID NO.3.
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CN115040695A (en) * 2021-03-09 2022-09-13 南开大学 Application of fusion protein active interface based on VE-cad-Fc/N-cad-Fc
CN115040695B (en) * 2021-03-09 2023-10-13 南开大学 Application of VE-cad-Fc/N-cad-Fc based fusion protein active interface
CN114213521A (en) * 2021-12-16 2022-03-22 中国海洋大学 Novel cell matrix-like biomaterial with wet adhesion performance and application thereof
CN114891818A (en) * 2022-06-13 2022-08-12 重庆大学 CD47-VE-M fusion protein, preparation method and application thereof
CN114891818B (en) * 2022-06-13 2023-09-01 重庆大学 CD47-VE-M fusion protein, preparation method and application thereof
CN117247441A (en) * 2023-11-16 2023-12-19 北京未名拾光生物技术有限公司 Recombinant mussel mucin and expression system and application thereof
CN117247441B (en) * 2023-11-16 2024-02-02 北京未名拾光生物技术有限公司 Recombinant mussel mucin and expression system and application thereof

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