CN105349546A - Apostichopus japonicus lectin gene, recombinant fusion protein and preparation method - Google Patents
Apostichopus japonicus lectin gene, recombinant fusion protein and preparation method Download PDFInfo
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- CN105349546A CN105349546A CN201510704039.6A CN201510704039A CN105349546A CN 105349546 A CN105349546 A CN 105349546A CN 201510704039 A CN201510704039 A CN 201510704039A CN 105349546 A CN105349546 A CN 105349546A
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Abstract
The invention discloses an Apostichopus japonicus lectin gene. A nucleotide sequence of the gene is shown as SEQ ID NO:1, and an amino acid sequence of recombinant fusion protein of the gene is shown as SEQ ID NO:2. The preparation method of the gene includes: extracting Apostichopus japonicus body wall total RNA; synthesizing a first strand of cDNA; using cDNA as a template to respectively obtain 5'RACE and 3'RACE products; using a RT-PCR primer for amplification to obtain a complete lectin gene segment, and obtaining a complete lectin sequence through sequencing and splicing; connecting the lectin gene onto a PMD19-T vector, and subjecting a product to positive colon expression and purification to obtain the Apostichopus japonicus lectin recombinant fusion protein rAj-MBCL. The preparation method is simple and high in purification rate, and the obtained Apostichopus japonicus lectin recombinant fusion protein is low in cost, high in agglutinating activity, high in expression quantity, small in experiment repeated difference, stable in result and high in controllability.
Description
Technical field
The invention belongs to marine organisms genetically engineered field, particularly relate to a kind of imitative stichopus japonicus agglutinin gene, recombination fusion protein and preparation method.
Background technology
Sea cucumber belongs to echinodermss Holothuroidea, about has kind more than 1200, and what be distributed in China marine site has kind more than 140, what wherein performance optimal, pharmaceutical use were the highest is originate in the ground such as LiaoNing, China, Shandong, Hebei the coastal imitative stichopus japonicus in North Pacific (
apostichopusjaponicus).In the last few years, sea cucumber had become the important ecological economy species in East Asia and south east asia, and in NORTH CHINA coastland, holothruian cultures industry also develops rapidly.But, owing to containing the disease problem that high density pathogenic bacteria causes holothruian cultures serious in ocean, constrain the sound development of holothruian cultures industry.The infringement of resisting foreign pathogen by improving sea cucumber natural immunity is the most scientific effective approach of sea cucumber healthy aquaculture.
There is the panimmunity factor in the immunity system of sea cucumber, comprise lectin, antibacterial peptide, N,O-Diacetylmuramidase, pattern recognition receptors and Complement C_3 etc., the research at present for lectin (Lectin) is the most extensive, thorough.Lectin is the nonimmune origin of a class, non-enzymatic, the protein that can impel cell agglutination or multivalence carbohydrate-binding protein, in its molecule containing one or more can with the non-catalytic structural domain of monose or the special Reversible binding of oligosaccharides, have sugar, glycolipid, glycoprotein height be affine and the characteristic of single-minded combination.Lectin has the function identifying cell surface specific carbohydrate structures, and participates in the vital movements such as apoptosis, cell adhesion, mitogenesis, transmembrane signal transduction and tumor cell.The reports such as Sun Yongxin, the lectin in sea cucumber body fluid has the effect of repairing wound; Play aggegation by combining with the specific carbohydrate structures on foreign cell and nurse one's health lethal effect.The research such as Cheung shows, the lectin separated in marine species has the various active such as efficient antibacterium, antimycotic, antiviral, anti-inflammatory, anti-parasitic and anticancer change, and visible agglutination element has vital role in the congenital immunity of marine invertebrate.But separation and Extraction natural sea cucumber lectin process is complicated, cost is high, up to now also not about the relevant report of restructuring imitative stichopus japonicus agglutinin gene, proteins encoded and preparation method.
Summary of the invention
The present invention is the above-mentioned technical problem in order to solve existing for prior art, provides a kind of imitative stichopus japonicus agglutinin gene, recombination fusion protein and preparation method.
Technical solution of the present invention is: a kind of imitative stichopus japonicus agglutinin gene, is characterized in that nucleotide sequence is as shown in SEQIDNO:1.
A recombination fusion protein for imitative stichopus japonicus agglutinin gene as above-mentioned in claim 1, is characterized in that aminoacid sequence is as shown in SEQIDNO:2.
A preparation method for above-mentioned imitative stichopus japonicus agglutinin gene, is characterized in that carrying out as follows successively:
A. imitative stichopus japonicus total serum IgE is extracted;
B. stichopus japonicus total serum IgE reverse transcription synthesis 3 '-RACE-ReadycDNA and 5 '-RACE-ReadycDNA will be imitated;
C. with 3 '-RACE-ReadycDNA for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-3 and UPM, obtain 3 '-FirstroundPCR product, described Auele Specific Primer Ajmbcl-3 sequence is as follows:
Ajmbcl-3：5'GGACTGGCTTCAATGGAAAGTGTTA3'；
D. with 3 '-FirstroundPCR product for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-n3 and NUP, obtain 3 '-NET-PCR product, described Auele Specific Primer Ajmbcl-n3 sequence is as follows:
Ajmbcl-n3：5'ACAGTCGCGGGGCCATAGCATCGGG3'；
E. with 5 '-RACE-ReadycDNA for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-5 and UPM, obtain 5 '-FirstroundPCR product, described Auele Specific Primer Ajmbcl-5 sequence is as follows:
Ajmbcl-5：5'GATGTACACCTGTGGATCCCAACCG3'；
F. with 5 '-FirstroundPCR product for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-n5 and NUP, obtain 5 '-NET-PCR product, described Auele Specific Primer Ajmbcl-n5 sequence is as follows:
Ajmbcl-n5：5'ATGACCACTGGCAAGAACATTACCT3'；
G. 3 '-NET-PCR product and the splicing of 5 '-NET-PCR product are obtained complete lectin gene segment.
The present invention with obtained complete lectin gene segment for template, complete agglutinin gene fragment is amplified by RT-PCR primer, and being connected on PMD19-T carrier, gained positive colony is expressed, purifying, namely obtains imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL.Preparation method is simple, purifying rate is high, the imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL cost obtained is low, agglutination activity is high, expression amount is many, experiment repetition difference is little, result stable, controllability is strong, can be used for the industrialized developing of sea cucumber immunostimulant, effectively can suppress sea cucumber disease problem, decreasing pollution, sets up healthy breeding environment.
Accompanying drawing explanation
Fig. 1 embodiment of the present invention imitates the agarose gel electrophoresis detection figure of stichopus japonicus lectin goal gene.
The double digestion electroresis appraisal figure of Fig. 2 embodiment of the present invention recombinant expression plasmid.
The SDS-PAGE gel electrophoresis of Fig. 3 embodiment of the present invention recombinant expression system (pET32a-AjMBCL) expression product and Westernblot qualification result schematic diagram.
Fig. 4 embodiment of the present invention imitates stichopus japonicus lectin recombination fusion protein rAj-MBCL blood coagulation activity analytical results schematic diagram.
Embodiment
A. imitative stichopus japonicus total serum IgE is extracted:
Get apostichopus japonicus body-wall 100mg, by liquid nitrogen grinding, add 1mlTRNzol – A
+extract reagent (TIANGEN), all the other concrete operations are according to the TRNzol-A of TIANGEN company
+total RNA extraction reagent specification sheets enters.
B. stichopus japonicus total serum IgE reverse transcription synthesis 3 '-RACE-ReadycDNA and 5 '-RACE-ReadycDNA will be imitated.
C. with 3 '-RACE-ReadycDNA for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-3 and UPM, obtain 3 '-FirstroundPCR product, described Auele Specific Primer Ajmbcl-3 sequence is as follows:
Ajmbcl-3：5'GGACTGGCTTCAATGGAAAGTGTTA3'。
D. with 3 '-FirstroundPCR product for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-n3 and NUP, obtain 3 '-NET-PCR product, described Auele Specific Primer Ajmbcl-n3 sequence is as follows:
Ajmbcl-n3：5'ACAGTCGCGGGGCCATAGCATCGGG3'；
The acquisition of 3 ' terminal nucleotide sequence: by 3 '-NET-PCR amplified production EasyPureQuickGel
Be connected with pMD19-T carrier after ExtractionKit purifying, send biological raw work order-checking, obtain 3 ' terminal sequence of imitative stichopus japonicus lectin Aj-MBCL gene.
E. with 5 '-RACE-ReadycDNA for template, carry out PCR with Auele Specific Primer Ajmbcl-5 and UPM
Reaction, obtain 5 '-FirstroundPCR product, described Auele Specific Primer Ajmbcl-5 sequence is as follows:
Ajmbcl-5：5'GATGTACACCTGTGGATCCCAACCG3'；
F. with 5 '-FirstroundPCR product for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-n5 and NUP, obtain 5 '-NET-PCR product, described Auele Specific Primer Ajmbcl-n5 sequence is as follows:
Ajmbcl-n5：5'ATGACCACTGGCAAGAACATTACCT3'；
The acquisition of 5 ' terminal nucleotide sequence: be connected with pMD19-T carrier after being purified with EasyPureQuickGelExtractionKit by 5 '-NET-PCR amplified production, send biological raw work order-checking, obtains 5 ' terminal sequence of imitative stichopus japonicus lectin Aj-MBCL gene.
G. 3 '-NET-PCR product and the splicing of 5 '-NET-PCR product are obtained complete lectin gene segment:
5 ' terminal sequence Seqman software of the imitative stichopus japonicus lectin Aj-MBCL gene that 3 ' terminal sequence of the stichopus japonicus lectin Aj-MBCL gene obtained by 3 '-RACE and 5 '-RACE obtain splices, and obtains complete imitative stichopus japonicus lectin Aj-MBCL gene nucleotide series.Through order-checking, nucleotide sequence is as shown in SEQIDNO:1, and the complete ORF contained, long is 537bp.
One. gene amplification, clone and the structure of recombinant plasmid:
1. with complete agglutinin gene fragment for template, carry out selective amplification by RT-PCR primer, described RT-PCR primer is as follows:
mAjmbcF:5'CCGGAATTCTGTACTTTGACGGCTTGTC3'EcoRI
mAjmbcR:5'CCCAAGCTTATTAAATTGATACACGGTG3'HindIII；
By RT-PCR product after detected through gel electrophoresis with EasyPureQuickGelExtractionKit carry out reclaiming, purifying.Agarose gel electrophoresis analysis chart is as shown in Figure 1: in Fig. 1, M:Marker(is followed successively by 2000,1000,750,500,250,100bp from top to bottom); 1:Aj-MBCL gene PCR amplified fragments, 477bp.
2. use the RT-PCR product of EcoR I and Hind III pair of purifying and expression vector pET-32a(+ again) carry out double digestion, electrophoresis reclaims object band, with T4ligase(TaKaRa) connect, transform BL21(DE3) competent cell, coating LB(AMP
+) agar plate, PCR identifies positive colony.PCR identifies that correct person is gene engineering expression bacterial strain, and restriction enzyme digestion and electrophoresis qualification as shown in Figure 2.
In Fig. 2, M:Marker(is followed successively by 8000,5000,3000,1500,1000,500bp from top to bottom);
1: recombinant expression plasmid (pET32a-Aj-MBCL); 2: recombinant expression plasmid (pET32a-Aj-MBCL) is through double digestion.
Above PCR reaction buffer, dNTP and enzyme are provided by TaKaRaLAPCR KitVer.2.1 test kit and by specification operates.
Two. the imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL of preparation
1. the expression of imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL:
The bacterium being accredited as positive colony is inoculated in LB(AMP
+) in liquid nutrient medium, 37 DEG C, 200 revs/min, activate 6 hours, be then inoculated in LB(AMP in the ratio of 1:100
+) liquid nutrient medium, 37 DEG C 200 revs/min are shaken bacterium 2.5 hours, to OD
600adding inducer isopropylthio thiogalactoside (IPTG) to final concentration when being 0.4 is 0.5mM, 28 DEG C, 80 revs/min of incubated overnight, collects thalline; 7000r/min, centrifugal 10min collects thalline, abandons supernatant, and raffinate is flowed out as far as possible, adds the ice-cold BingdingBuffer(10mM imidazoles of 40ml with the nutrient solution of every 100ml; 0.5M sodium-chlor; 20mMTris-HCl, PH7.9) proportion re-suspended cell; Be placed in ultrasonic disruption lysing cell on ice, intensity 50%, 10min, pulseon10s, pulseoff5s, 5000r/min, collecting precipitation.This protein expression is inclusion body, and the SDS-PAGE gel electrophoresis of protein expressioning product and Westernblot analytical results are as shown in Figure 3.
A figure in Fig. 3 is SDS-PAGE gel electrophoresis analysis figure, M is that albumen Marker(is followed successively by 200,116,97.2,66.4,44.3,29.0,20.1,14.3,6.5KD from top to bottom) 1:pET32a/BL21 expression product do not induce; 2:pET32a/BL21 expression product is through induction; 3:pET32a-Aj-MBCL/BL21 fusion expressed product is not induced; 4:pET32a-Aj-MBCL/BL21 fusion expressed product is through induction; The product of 5:pET32a-Aj-MBCL fusion rotein after affinitive layer purification.
B figure is Westernblot analysis chart, 1: be BSA contrast; 2: fusion rotein dilutes 20 times; 3: fusion rotein dilutes 10 times.
2. the separation of imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL:
Adopt the method purification of Recombinant fusion rotein of affinity chromatography.
The disposable sterilization device of the solution of obtained inclusion body after sex change, renaturation with 0.45 μm is filtered; Gel is filled with post (ProteinIso
tMni-NTAResin, transgen), use the deionized water of 3 times of volumes and the BingdingBuffer(10mM imidazoles of 8 times of volumes successively; 0.5M sodium-chlor; 20mMTris-HCl, PH7.9) balance, flow velocity is 2ml/min; By cell pyrolysis liquid upper prop, flow velocity is 2ml/min; Use the BingdingBuffer of 15 times of volumes to rinse pillar, then use the WashingBuffer(60mM imidazoles of 5 times of volumes; 0.5M sodium-chlor; 20mMTris-HCl, PH7.9) rinse pillar, finally use the ElutionBuffer(0.5M imidazoles of 3 times of volumes; 0.5M sodium-chlor; 20mMTris-HCl, PH7.9) wash-out, obtain purer imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL.
The Denaturation and Renaturation concrete steps of inclusion body are as follows:
By inclusion body by 1:10(W/V) be resuspended in washings (20mMTris-HCl, PH7.9; 1.5mM urea; Dispel 1mMEDTA), the centrifugal 30min of 1000rpm; , the inclusion body washed is added denaturation buffer (20mMTris-HCl, PH7.9 according to the ratio of 1:15; 8M urea; 1mMEDTA), room temperature places 1-2 hour; By centrifugal for solution 12000rpm 30min, get supernatant; The albumen that sex change is good puts into dialysis tubing, is immersed in renaturation buffer (20mMTris-HCl, PH7.9) to stir to spend the night, and obtains activated albumen.
Measure the imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL concentration of purifying by BCA method, operate and carry out according to BCA determination of protein concentration test kit (the green skies) specification sheets.
Check order to obtained imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL, its aminoacid sequence is 178 amino acid whose polypeptide as shown in SEQIDNO:2, and this polypeptide theoretical protein matter molecular weight is 17.59KD.
Three. imitative stichopus japonicus lectin recombination fusion protein rAj-MBCL detects HRBC agglutination activity:
1. erythrocytic process
Get human blood 5ml, with 3.8%(M/M) liquor sodii citratis anti-freezing, by anticoagulation through the centrifugal 10min of 2000rmp, removing blood plasma.Add the TBS solution of 10 times of volumes, fully wash, the centrifugal 5min of 1000rmp removes supernatant, repeats 4 times.With TBS-Ca
2+solution is made into 2%(V/V) red cell suspension, be simultaneously made into 2%(V/V with TBS solution) red cell suspension; Separately be made into 2%(V/V with TBS-EGTA) red cell suspension.
2. experimental design:
In each hole, add corresponding damping fluid (each 80 μ l), A group is TBSBuffer; B, D group is TBSBuffer+Ca
2+; C group is TBSBuffer+Ca
2++ EDTA.A, B, C group first hole all adds 80 μ lAjMBCL albumen (0.24mg/ml); The BSA that D group adds same concentrations contrasts.Get 80 μ l mixed solutions after mixing and add second hole, get 80 μ l after mixing and add the 3rd hole, do doubling dilution by that analogy, 1:2(0.12), 1:4(0.06), 1:8(0.03), 1:16(0.015), 1:32(0.0075), 1:64(0.0038), 1:128(0.0019), 1:236(0.00095).Add 5% red corpuscle 20 μ l in the air each, after mixing, room temperature leaves standstill 5min, and scanner qualification result as shown in Figure 4.
In Fig. 4:
Stichopus japonicus lectin recombination fusion protein rA-jMBCL and TBSbuffer compound sample are imitated in A behavior;
B behavior is imitated stichopus japonicus lectin recombination fusion protein rA-jMBCL and TBSbuffer(and is contained 20mMCaCl
2) compound sample;
C behavior is imitated stichopus japonicus lectin recombination fusion protein rA-jMBCL and TBSbuffer(and is contained 20mMEDTA and 20mMCaCl
2) compound sample;
D behavior bovine serum albumin BSA and TBSbuffer(contains 20mMCaCl
2) compound sample;
1-8 is classified as fusion rotein to be measured and BSA by the concentration of doubling dilution, is followed successively by 0.24,0.12,0.06,0.03,0.015,0.0075,0.0038, and 0.0019mg/ml.
3. result:
Result shows, and the red corpuscle of A, C, D group is dispersed; And B group has obvious agglutination phenomenon, reduce gradually, 1,2 from 1 to 8 agglutination activities, No. 3 agglutination activities are comparatively obvious, and 4, No. 5 have faint agglutination activity, show activity of lectin hardly after No. 6.Reach a conclusion from result, imitative stichopus japonicus lectin recombination fusion protein rA-jMBCL of the present invention is Ca
2+dependent form, has obvious agglutination phenomenon when the metal ion chelation agents such as EDTA have suppression to be greater than 0.03mg/ml as concentration to it.
Sequence table
<110> Dalian Ocean University
<120> imitates stichopus japonicus agglutinin gene, recombination fusion protein and preparation method
<160>8
<210>1
<211>537
<212>DNA
<213> imitates stichopus japonicus agglutinin gene
<400>1
1ATGTATCGCACCGTTTTTCTGCTCGTTGTCACTTGCATGTTGTTTGTGCATTCCAGCGGC
61TGTACTTTGACGGCTTGTCCATCAAAATGGACTGGCTTCAATGGAAAGTGTTACAGATTG
121TTTGCTGCTGGACACAAACAATTTGATGCCGCGGAGAGAGCCTGTCAAAGTGCAAAACTC
181GTTGACTGCCAAGGTAATGTTCTTGCCAGTGGTCATCTGGCATCTGTTCACAGTCAAGAA
241GAACAAAACTTTCTCTTAGAAATGGTTCGGTCGACTTTACAATATACAAGCGGTTGGGAT
301CCACAGGTGTACATCGGAATGAAAGTTGGATACCACAACAACCAGCAAAGTTGGACCGAT
361GGATCATCCGTTGATTACACCAGCTGGTTCTCTGGAGAGCCGAACAATGGGCCTAACAGT
421CGCGGGGCCATAGCATCGGGCTTGCATTCGCGTGGTAAATGGGCCGATGTATACAGCAAC
481AGCAACTTCCCCTATATTTGTCAGCTACCTTGCACCGATTATCAATTTAATTGGTAG537
<210>2
<211>179
<212>PRT
<213> imitates the recombination fusion protein of stichopus japonicus agglutinin gene
<400>2
1MYRTVFLLVVTCMLFVHSSG
21CTLTACPSKWTGFNGKCYRL
41FAAGHKQFDAAERACQSAKL
61VDCQGNVLASGHLASVHSQE
81EQNFLLEMVRSTLQYTSGWD
101PQVYIGMKVGYHNNQQSWTD
121GSSVDYTSWFSGEPNNGPNS
141RGAIASGLHSRGKWADVYSN
161SNFPYICQLPCTDYQFNW*178
<210>3
<211>25
<212>DNA
<213> artificial sequence
<220>
<223> primer
<400>3
GGACTGGCTTCAATGGAAAGTGTTA25
<210>4
<211>25
<212>DNA
<213> artificial sequence
<220>
<223> primer
<400>4
ACAGTCGCGGGGCCATAGCATCGGG25
<210>5
<211>25
<212>DNA
<213> artificial sequence
<220>
<223> primer
<400>5
GATGTACACCTGTGGATCCCAACCG25
<210>6
<211>25
<212>DNA
<213> artificial sequence
<220>
<223> primer
<400>6
ATGACCACTGGCAAGAACATTACCT25
<210>7
<211>28
<212>DNA
<213> artificial sequence
<220>
<223> primer
<400>7
CCGGAATTCTGTACTTTGACGGCTTGTC28
<210>8
<211>28
<212>DNA
<213> artificial sequence
<220>
<223> primer
<400>5
CCCAAGCTTATTAAATTGATACACGGTG28
Claims (3)
1. an imitative stichopus japonicus agglutinin gene, is characterized in that nucleotide sequence is as shown in SEQIDNO:1.
2. imitate a recombination fusion protein for stichopus japonicus agglutinin gene as claimed in claim 1, it is characterized in that aminoacid sequence is as shown in SEQIDNO:2.
3. imitate a preparation method for stichopus japonicus agglutinin gene as claimed in claim 1, it is characterized in that carrying out as follows successively:
A. imitative stichopus japonicus total serum IgE is extracted;
B. stichopus japonicus total serum IgE reverse transcription synthesis 3 '-RACE-ReadycDNA and 5 '-RACE-ReadycDNA will be imitated;
C. with 3 '-RACE-ReadycDNA for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-3 and UPM, obtain 3 '-FirstroundPCR product, described Auele Specific Primer Ajmbcl-3 sequence is as follows:
Ajmbcl-3：5'GGACTGGCTTCAATGGAAAGTGTTA3'；
D. with 3 '-FirstroundPCR product for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-n3 and NUP, obtain 3 '-NET-PCR product, described Auele Specific Primer Ajmbcl-n3 sequence is as follows:
Ajmbcl-n3：5'ACAGTCGCGGGGCCATAGCATCGGG3'；
E. with 5 '-RACE-ReadycDNA for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-5 and UPM, obtain 5 '-FirstroundPCR product, described Auele Specific Primer Ajmbcl-5 sequence is as follows:
Ajmbcl-5：5'GATGTACACCTGTGGATCCCAACCG3'；
F. with 5 '-FirstroundPCR product for template, carry out PCR reaction with Auele Specific Primer Ajmbcl-n5 and NUP, obtain 3 '-NET-PCR product, described Auele Specific Primer Ajmbcl-n5 sequence is as follows:
Ajmbcl-n5：5'ATGACCACTGGCAAGAACATTACCT3'；
G. 3 '-NET-PCR product and the splicing of 5 '-NET-PCR product are obtained complete lectin gene segment.
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| CN106666064A (en) * | 2016-12-24 | 2017-05-17 | 山东明鑫集团有限公司 | Preparation method of sea cucumber-like protein |
| CN108191964A (en) * | 2018-01-11 | 2018-06-22 | 大连海洋大学 | Imitative stichopus japonicus F type agglutinins AjFL-1, preparation method and application |
| CN109628460A (en) * | 2019-01-18 | 2019-04-16 | 自然资源部第三海洋研究所 | Derived antimicrobial peptide hydramacin and preparation method thereof is carried out in a kind of hadal |
| CN114891084A (en) * | 2022-06-01 | 2022-08-12 | 辽宁省海洋水产科学研究院 | Application of gene recombinant sea cucumber peptide rAj-HRP in antitumor drugs |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106666064A (en) * | 2016-12-24 | 2017-05-17 | 山东明鑫集团有限公司 | Preparation method of sea cucumber-like protein |
| CN108191964A (en) * | 2018-01-11 | 2018-06-22 | 大连海洋大学 | Imitative stichopus japonicus F type agglutinins AjFL-1, preparation method and application |
| CN108191964B (en) * | 2018-01-11 | 2021-05-11 | 大连海洋大学 | Apostichopus japonicus F-type lectin AjFL-1, and preparation method and application thereof |
| CN109628460A (en) * | 2019-01-18 | 2019-04-16 | 自然资源部第三海洋研究所 | Derived antimicrobial peptide hydramacin and preparation method thereof is carried out in a kind of hadal |
| CN114891084A (en) * | 2022-06-01 | 2022-08-12 | 辽宁省海洋水产科学研究院 | Application of gene recombinant sea cucumber peptide rAj-HRP in antitumor drugs |
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Application publication date: 20160224 |