CN107987154A - Long oyster IgSF molecule CgCAICP1 gene recombinant proteins, preparation method and application - Google Patents

Long oyster IgSF molecule CgCAICP1 gene recombinant proteins, preparation method and application Download PDF

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Publication number
CN107987154A
CN107987154A CN201711421622.1A CN201711421622A CN107987154A CN 107987154 A CN107987154 A CN 107987154A CN 201711421622 A CN201711421622 A CN 201711421622A CN 107987154 A CN107987154 A CN 107987154A
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cgcaicp1
long oyster
recombinant protein
seq
gene recombinant
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CN107987154B (en
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宋林生
刘冬杨
衣启麟
宋小瑞
王玲玲
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Dalian Ocean University
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Dalian Ocean University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Abstract

The present invention discloses a kind of long oyster IgSF moleculesCgCAICP1Gene recombinant protein, amino acid sequence is as shown in SEQ ID NO.1.Preparation method carries out in accordance with the following steps successively：With specific primer P1 and P2 to long oyster IgSF moleculesCgCAICP1Coding domain segment carries out PCR amplification, and the DNA sequence dna of the primer P1 is as shown in SEQ ID NO.2, and the DNA sequence dna of the primer P2 is as shown in SEQ ID NO.3；Pcr amplification product and pET32a carriers are passed throughNcoI andHindConnected, converted, sequencing identification recon by T4 ligases after III digestion；Above-mentioned recon is transferred to Escherichia coli Transetta（DE3）Fiber differentiation is carried out in expression bacterial strain, then purifying, renaturation.The long oyster IgSF moleculesCgCAICP1Gene recombinant protein, which can be applied to prepare, suppresses Gram negative bacteria drugs.

Description

Long oyster IgSF moleculesCgCAICP1Gene recombinant protein, preparation method and application
Technical field
The invention belongs to technical field of molecular biology, more particularly to a kind of long oyster IgSF moleculesCgCAICP1Gene weight Histone, preparation method and application.
Background technology
Immunoglobulin superfamily IgSF molecules, are mainly produced by bone-marrow-derived lymphocyte, are vertebrate adaptive immunities （adaptive immunity）Important composition component, a series of molecular mechanisms can be passed through and produce diversified antigen binding Epitope, and combined with corresponding epitopic specificity.Although invertebrate does not possess in vertebrate based on antibody Adaptive immune system, but it still is able to form effective immune response for cause of disease invasion.Research finds, IgSF molecule rows Make the major way of function include following four type：A. pattern-recognition；B. microorganism agglutination；C. opsonic action is swallowed； D. cell agglutination and anti-agglutination.As the hemolin in insect can be combined with the two basic change site on LPS and can Aggegation multiple-microorganism；The DSCAM in VCBP and crustacean in lancelet can also be identified and be combined a large amount of ligands；In The bright prawn FcLec4 of state is combined with the β-integrin on blood lymphocyte surface and is played opsonic action.
But so far not on long oyster IgSF moleculesCgCAICP1Gene recombinant protein, preparation method and The relevant report applied in suppression Gram negative bacteria drugs are prepared.
The content of the invention
The present invention is to solve the above-mentioned technical problem present in the prior art, there is provided a kind of long oyster IgSF moleculesCgCAICP1Gene recombinant protein, preparation method and application.
The present invention technical solution be：A kind of long oyster IgSF moleculesCgCAICP1Gene recombinant protein, its feature It is：Amino acid sequence is as shown in SEQ ID NO.1.
Above-mentioned long oyster IgSF moleculesCgCAICP1The preparation method of gene recombinant protein, it is characterised in that successively according to such as Lower step carries out：
A. with specific primer P1 and P2 to long oyster IgSF moleculesCgCAICP1Coding domain segment carries out PCR amplification, the primer The DNA sequence dna of P1 is as shown in SEQ ID NO.2, and the DNA sequence dna of the primer P2 is as shown in SEQ ID NO.3；
B. pcr amplification product and pET32a carriers are passed throughNcoI andHindConnected, converted by T4 ligases after III digestion, surveyed Sequence identifies recon；
C. above-mentioned recon is transferred to Escherichia coli Transetta（DE3）Fiber differentiation is carried out in expression bacterial strain, it is then pure Change, renaturation, that is, obtain the recombinant protein with amino acid sequence in sequence table SEQ ID NO.1.
Above-mentioned long oyster IgSF moleculesCgCAICP1Gene recombinant protein is in suppression Gram negative bacteria drugs are prepared Using.
Long oyster IgSF molecules in the present inventionCgCAICP1Gene recombinant protein is cloned from long oyster cDNA library, is had The combination of multiple-microorganism is active and can significantly improve phagocytic rate of the long oyster blood lymphocyte for Gram-negative bacteria, As a kind of effective pattern recognition receptors, antimicrobial DP finish, novel immune preparation and feed addictive etc. are being prepared With application value.
Brief description of the drawings
Fig. 1 is IgSF molecules of the embodiment of the present inventionCgCAICP1The bacterium of gene recombinant protein combines Activity determination figure.
Fig. 2 is IgSF molecules of the embodiment of the present inventionCgCAICP1Gene recombinant protein promotees phagocytosis detection result figure.
Embodiment
The above-mentioned long oyster IgSF molecules of the present inventionCgCAICP1The preparation method of gene recombinant protein,
Carry out in accordance with the following steps successively：
1. the structure of recombinant vector
The recombinant vector that the embodiment of the present invention uses for Novagen companies pET-32a (+) prokaryotic expression carrier.Pass through PCR skills Art, with the addition of respectively using 5 ' endsNcoI andHind Primer P1 and the P2 amplification of restriction enzyme site come from long oysterCgCAICP1 The code area of mRNA.The DNA sequence dna of the primer P1 is as shown in SEQ ID NO.2, the DNA sequences of the primer P2 Row are as shown in SEQ ID NO.3.
PCR reaction conditions are：94 DEG C of 5 min of pre-degeneration first, subsequently into following circulation：94 DEG C be denatured 30 seconds, 56 DEG C annealing 30 seconds, 72 DEG C extension 2 min, altogether carry out 30 circulation, it is last 72 DEG C extension 10 min.It will be expanded with agarose gel electrophoresis Increase fragment purification recycling, be connected with pMD19-T carriers.Screening positive clone after conversion, extracts plasmid, and usesNcoI andHind Double digestion is carried out to plasmid；Purpose fragment is recycled, and is passed throughNcoI andHind The expression vector pET-32a (+) of digestion is even Connect, complete the structure of recombinant plasmid.
2. recombinant protein rCgThe expression of CAICP1
By in the recombinant plasmid transformed expressive host bacterium Escherichia coli Transetta (DE3) built, picking monoclonal, is inoculated with In the LB fluid nutrient mediums of 200 ml, 220 rpm, 37 DEG C of cultures to OD 600= 0.4~0.8.Add IPTG（Final concentration 1 mmol L-1）, continue after 4 DEG C of 10000 rpm to centrifuge 5 min when culture 4 is small, collect thalline, frozen in -80 DEG C spare； Take 1 ml bacterium solutions to centrifuge at the same time, after supernatant discarding, add 80 μ l water and 5 × albumen sample-loading buffer of 20 μ l, 99 DEG C are boiled 10 min are boiled, are slightly centrifuged, SDS-PAGE detection expression products.
3. recombinant protein rCgThe purifying of CAICP1 and renaturation
Recombinant protein uses nickel Ago-Gel FF column purifications, denaturation recombinant protein is obtained, with elution buffer dialysis renaturation. Concrete operation step is as follows：
（1）Nickel Ago-Gel FF fills column, and 1.6 × 20cm, bed volume is 10 ml；
（2）With buffer solution I（50 mmol L-1Tris-Hcl buffer solutions, pH=7.4,50 m mol L-1 NaCl, 8 mol L-1Urea）2 ~ 5 bed volumes are balanced, flow velocity is 2 ml min-1；
（3）The cell for IPTG induced expressions of learning from else's experience, is resuspended, 150 W ultrasonications 30 min, 12000 rmp, 4 with buffer solution I DEG C centrifugation 30 min, supernatant use after 0.45 μm of membrane filtration, mistake column, flow velocity is 1 ml min-1；
（4）2 ~ 5 bed volumes are washed again with buffer solution 1, and flow velocity is 2 ml min-1；
（5）With there is 50 mmol L-1The buffer solution I of imidazoles washes 2 ~ 5 bed volumes again, and flow velocity is 2 ml min-1；
（6）With 400 mmol L-1The buffer solution I elution destination proteins of imidazoles, are collected;
（7）With the expression of SDS-PAGE detection fusion albumen；
（8）5 bed volumes are washed with stream of pure water, then 3 bed volumes are washed with 20% ethanol stream, flow velocity is 2 ml min-1, column Son, which is placed in the recombinant protein for preserving in 4 DEG C of environment and being purified under denatured state, to be needed to remove urine by dialysing in renaturation buffer Element, makes albumen correctly fold again, recovers correct conformation.It is denatured reduced glutathione of the purified product through 2 mM, 0.4 mM oxygen Change what glutathione, 1 mM EDTA, 50 mM Tris-HCl, 100 mM NaCl, 10% glycerine, 1% glycine and gradient reduced Urea dialysis renaturation, urea concentration are gradually substituted into 4 M, 3 M, 2 M, 1M, 0 M, last time dialysis to nothing from 6 M of starting Not glycerol adding during the dialyzate of urea, dialyse 12 h at 4 DEG C every time.Obtain long oysterCgCAICP1 gene recombinant proteins （rCgCAICP1）.
Obtained long oysterCgCAICP1Gene recombinant protein amino acid sequence is as shown in SEQ ID NO.1.
Length：574 amino acid
Type：Amino acid
Chain：It is single-stranded
Characteristic：Molecular weight is 63.4 kDa, and isoelectric point 5.91, has cysteine rich domain domain and three immune globulins White domain.
Experimental example 1：Long oyster recombinant protein rCgCAICP1 bacterium combine Activity determination
Recombinant protein is detected to two kinds of Gram-negative bacterias based on western blotting methods（Vibrio splindidus and large intestine bar Bacterium）, two kinds of gram-positive bacterias（Staphylococcus aureus and micrococcus luteus）And a kind of combination of fungi Pichia pastoris is lived Property.Strain source used is as follows：Vibrio splindidus（Vibrio splendidus JZ6）Purchased from the Microbiological Culture Collection of Beijing The heart, Escherichia coli（Escherichia coli）Purchased from Beijing Quan Shi King Companies, staphylococcus aureus（Staphylococcus aureus）Purchased from Beijing Culture Collection, micrococcus luteus（Micrococcus luteus）Purchased from the micro- life in Beijing Thing Culture Collection Center, Pichia yeast（Pichia pastoris GS115）Purchased from Invitrogen companies.
Concrete operations are as follows：
（1）Above-mentioned 5 kinds of microorganisms are incubated overnight, cultural method：Micrococcus luteus and Escherichia coli are in LB culture mediums 37 When °C culture 20 is small, for staphylococcus aureus when 28 °C of LB culture mediums culture 20 is small, Vibrio splindidus is in 2216E culture mediums 28 °C culture 20 it is small when, Pichia yeast in YPD culture mediums 28 °C culture 20 it is small when；
（2）Thalline is collected by centrifugation, is resuspended using TBS buffer solutions, adjustment bacteria concentration is 1 × 108 CFU；
（3）Draw 100 μ l microorganisms suspensions and long oyster obtained by isometric embodiment of the present inventionCgCAICP1 genetic recombination eggs White mixing, rotates in room temperature and is incubated 30 min；
（4）10000 rpm centrifuge 2 min and collect thalline, TBS buffer solution for cleaning thalline four times；
（5）Thalline is collected after the completion of washing, is resuspended using 40 μ l sterile waters；
（6）5 × protein electrophoresis buffer solution of 10 μ l is added, 10 min, SDS-PAGE electrophoretic separation albumen samples are heated in 99 °C Product；
（7）After the completion of electrophoresis, gel is removed, and cuts the NC films of formed objects and filter paper immerses electricity and turns to stand in buffer solution jointly 10 min；
（8）Filter paper, NC films, gel and filter paper are put into electroporation successively according to order from top to bottom, according to gel piece area Size sets corresponding electric current, 65 min of transferring film；
（9）NC films are taken out, are washed three times using TBS buffer solutions, every time 5 min；
（10）Washed three times using buffer solution TBST, every time 5 min；
（11）NC films are put into 5% skimmed milk power（It is dissolved in TBST）In, when room temperature closing 2 is small；
（12）NC films are taken out, are washed three times using TBST buffer solutions, every time 5 min；
（13）NC films are immersed into diluted label monoclonal antibody solution in proportion（5% skimmed milk power, TBST buffer solutions）, in room temperature condition It is lower be incubated 1 it is small when；
（14）NC films are taken out, are washed three times using TBST buffer solutions, every time 5 min;
（15）NC films are put into diluted sheep anti-Mouse HRP secondary antibodies in proportion（Work is given birth to purchased from Shanghai）In solution（5% defatted milk Powder TBST buffer solutions）, when incubation at room temperature 1 is small；
（16）NC films are taken out, are washed three times using TBST buffer solutions, every time 10 min；
（17）ECL methods are developed, and imager records western blotting as a result, as shown in the figure.
The long oyster of the results show present inventionCgCAICP1 gene recombinant proteins and two kinds of grams and Staphylococcus aureus Bacterium shows stronger combination activity, and weaker with the binding ability of micrococcus luteus and Pichia pastoris.TRX negative controls and Appearance in the blank control of cleaning solution then without obvious band.
Experimental example 2：Long oyster recombinant protein rCgCAICP1 promotees phagocytic activity detection
Using fluorescein isothiocynate（FITC）To three kinds of microorganisms（Vibrio splindidus, micrococcus luteus, Pichia pastoris）Into rower Note, phagocytosis efficiency of the method then counted using fluorescence microscope to oyster blood lymphocyte are detected.
The strain source is as above.
Concrete operations are as follows：
（1）Above-mentioned three kinds of microorganisms are cultivated respectively and collect thalline；
（2）Mixed using formaldehyde with microorganism, fix 10 min；
（3）Bacterium is collected by centrifugation in 4000 × g, 10min.The NaHCO of 0.1 M3After washing three times, it is incubated in containing 0.1 mg/ml FITC's 0.1M NaHCO3In, 25 °C of 2 h of incubation, and with slight wobble；
（4）After supernatant is abandoned in centrifugation, using TBS buffer solutions by micro-organism washing to colourless；
（5）Thalline will be resuspended using TBS and adjust concentration as 108 CFU ml-1；
（6）After addition equal proportion anti-coagulants extracts hemolymph out, 4 °C, 800 × g, 10 min are centrifuged, collect haemocyte；
（7）Cell concentration is adjusted to 10 with L15 plus salt cell culture medium6 cells ml-1;
（8）By long oyster obtained by the embodiment of the present inventionCgCAICP1 gene recombinant proteins and cell are with 1:1 volume ratio, room temperature Lucifuge is incubated 30 min；
（9）Isometric bacterium marked is added, room temperature lucifuge is incubated 30 min；
（10）Drip piece, every 80 μ l of slice, thin piece；
（11）In wet box, room temperature settles 1 h；
（12）Incline residual liquid, and blots residual liquid around drop, with 2 mg ml-110 min of trypan blue quenching fluorescence；
（13）20 min are fixed in acetone, necessarily ensure that acetone did not had glass slide；
（14）PBS is washed three times, every time 5 min；
（15）DAPI dyes 3 min；
（16）PBS is washed three times；
（17）In fluorescence microscopy Microscopic observation, and phagocyte and non-phagocytic cell are counted, the results are shown in Figure 2.
The long oyster of the results show embodiment of the present inventionCgCAICP1Gene recombinant protein has Vibrio splindidus obvious rush to gulp down Effect is bitten, the phagocytosis to micrococcus luteus also has certain effect, and then promotees Phagocytosis without significant for Pichia pastoris.It is right According to group rTRX three kinds of microorganisms are not detected by with obvious rush Phagocytosis.
Sequence table
<110>Dalian Ocean University
<120>Long oyster IgSF molecule CgCAICP1 gene recombinant proteins, preparation method and application
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 574
<212> PRT
<213>Long oyster (Crassostrea gigas)
<400> 1
Met Ser Asp Lys Ile Ile His Leu Thr Asp Asp Ser Phe Asp Thr Asp
1 5 10 15
Val Leu Lys Ala Asp Gly Ala Ile Leu Val Asp Phe Trp Ala Glu Trp
20 25 30
Cys Gly Pro Cys Lys Met Ile Ala Pro Ile Leu Asp Glu Ile Ala Asp
35 40 45
Glu Tyr Gln Gly Lys Leu Thr Val Ala Lys Leu Asn Ile Asp Gln Asn
50 55 60
Pro Gly Thr Ala Pro Lys Tyr Gly Ile Arg Gly Ile Pro Thr Leu Leu
65 70 75 80
Leu Phe Lys Asn Gly Glu Val Ala Ala Thr Lys Val Gly Ala Leu Ser
85 90 95
Lys Gly Gln Leu Lys Glu Phe Leu Asp Ala Asn Leu Ala Gly Ser Gly
100 105 110
Ser Gly His Met His His His His His His Ser Ser Gly Leu Val Pro
115 120 125
Arg Gly Ser Gly Met Lys Glu Thr Ala Ala Ala Lys Phe Glu Arg Gln
130 135 140
His Met Asp Ser Pro Asp Leu Gly Thr Asp Asp Asp Asp Lys Ala Met
145 150 155 160
Ala Gln Asp Ser Ser Gln Met Pro Pro Asn Met Thr Ile Thr Phe Asn
165 170 175
Asn Glu Tyr Phe Phe Pro Arg Gly Ser Ile Leu Ala Asp Ala Ser Phe
180 185 190
Asn Cys Thr Ala Glu Asn Gly Arg Val Leu Tyr Glu Trp Lys Lys Asp
195 200 205
Gly Asn Val Val Gln Asn Thr Gln Ser Val Thr Val Asp Asn Ser Thr
210 215 220
Gly Ile Leu Lys Phe His Gln Met Gln Asn Gly Asp Tyr Gly Thr Tyr
225 230 235 240
Gln Cys Phe Ala Thr Asn Gly Tyr Gly Thr Ser Leu Ser Lys Pro Phe
245 250 255
Lys Ile Met Glu Ala Arg Leu Gly Ser Phe Pro Thr Ser Gln Thr Gln
260 265 270
Glu Ile Lys Cys Glu Glu Phe Lys His Cys Lys Val Glu Cys Arg Tyr
275 280 285
Lys Pro Thr Cys Leu Pro Glu Ser Gln Cys Lys Val Glu Trp Lys Ile
290 295 300
Gly Glu Gly Thr Lys Thr Asn Val Glu Ile Asn Lys Arg Val Gly Val
305 310 315 320
Asp Gly Asn Gly Gly Leu His Phe Leu Trp Thr Ser Met Ser Asp Trp
325 330 335
Thr Gly Gln Gln Tyr Arg Cys Gly Val Trp His Glu Gln Leu Lys Thr
340 345 350
Leu Val Val Gly Ser Gln Thr Ser Leu Lys Ile Asp Ser Ala Thr Ala
355 360 365
Val Pro Lys Val Asp Pro Met Leu Val Phe Lys Glu Asn Gly Lys Ala
370 375 380
Leu Ile Gly Glu Arg Gly Val Leu Arg Cys Met Phe Ser Gly Tyr Pro
385 390 395 400
Val Pro Asp Ile Thr Trp Ile Ser Pro Gln Lys Met Asn Ile Asp Asp
405 410 415
Ala Asp Gly Lys Tyr Glu Ile Ser Asp Phe Gly Arg Val Leu Thr Ile
420 425 430
Leu Arg Ala Glu Ser Lys His Glu Gly Thr Tyr Thr Cys Lys His Ser
435 440 445
Gly Lys Asn Glu Thr Val Phe Leu Asn Ala Thr Ser Ala Pro Phe Leu
450 455 460
Asn Gly Ser Asn Gln Met Gln Asp Leu Val Leu Pro Glu Gly Gln Glu
465 470 475 480
Ala Thr Phe Arg Cys Glu Ala Glu Ser Ser Pro Asp Glu Leu Pro Pro
485 490 495
Thr Arg Pro Thr Trp Lys Lys Asn Gly Val Asp Leu Lys Ile Asp Gly
500 505 510
Gly Lys Tyr Leu Leu Gly Glu Asn Ser Gln Val Leu Ser Leu Lys Asp
515 520 525
Val Gln Lys Ser Asp Ser Gly Val Tyr Gln Cys Met Ser Glu Asn Ser
530 535 540
Glu Gly Val Leu Leu Lys Glu Ala Ile Leu Lys Val Thr Asp Pro Ile
545 550 555 560
Lys Lys Leu Ala Ala Ala Leu Glu His His His His His His
565 570
<210> 2
<211> 34
<212> DNA
<213>Artificial sequence (Crassostrea gigas)
<400> 2
<210> 3
<211> 32
<212> DNA
<213>Artificial sequence (Crassostrea gigas)
<400> 3

Claims (3)

  1. A kind of 1. long oyster IgSF moleculesCgCAICP1Gene recombinant protein, it is characterised in that：Amino acid sequence such as SEQ ID Shown in NO.1.
  2. A kind of 2. long oyster IgSF molecules as claimed in claim 1CgCAICP1The preparation method of gene recombinant protein, its feature It is to carry out in accordance with the following steps successively：
    A. with specific primer P1 and P2 to long oyster IgSF moleculesCgCAICP1Coding domain segment carries out PCR amplification, the primer The DNA sequence dna of P1 is as shown in SEQ ID NO.2, and the DNA sequence dna of the primer P2 is as shown in SEQ ID NO.3；
    B. pcr amplification product and pET32a carriers are passed throughNcoI andHindConnected, converted by T4 ligases after III digestion, surveyed Sequence identifies recon；
    C. above-mentioned recon is transferred to Escherichia coli Transetta（DE3）Fiber differentiation is carried out in expression bacterial strain, it is then pure Change, renaturation, that is, obtain the recombinant protein with amino acid sequence in sequence table SEQ ID NO.1.
  3. A kind of 3. long oyster IgSF molecules as claimed in claim 1CgCAICP1Gene recombinant protein is preparing suppression gram-negative Application in property bacterium medicine.
CN201711421622.1A 2017-12-25 2017-12-25 Ostrea gigas IgSF molecule CgCAICP1 gene recombinant protein, preparation method and application Active CN107987154B (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113912691A (en) * 2021-11-01 2022-01-11 大连海洋大学 Recombinant crassostrea gigas high-mobility group protein r-CgHMGB1, preparation method and application thereof
CN114044816A (en) * 2021-11-10 2022-02-15 大连海洋大学 Recombinant crassostrea gigas pyrogallin protein rCgGSDME-N, preparation method and application thereof

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DONGYANG LIU等: "A hypervariable immunoglo bulin superfamily member from Crassostrea gigas functions as pattern recognition receptor with opsonic activity", 《DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY》 *
GENBANK DATABASE: "PREDICTED: Crassostrea gigas neurofascin (LOC1 05322290), transcript variant X1 , mRNA", 《GENBANK DATABASE》 *
刘聪辉: "长牡蛎免疫球蛋白超家族（IgSF）成员结构与功能的研究", 《中国博士学位论文全文数据库（电子期刊）农业科技辑》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113912691A (en) * 2021-11-01 2022-01-11 大连海洋大学 Recombinant crassostrea gigas high-mobility group protein r-CgHMGB1, preparation method and application thereof
CN113912691B (en) * 2021-11-01 2023-08-11 大连海洋大学 Recombinant crassostrea gigas high mobility group protein r-CgHMGB1, preparation method and application thereof
CN114044816A (en) * 2021-11-10 2022-02-15 大连海洋大学 Recombinant crassostrea gigas pyrogallin protein rCgGSDME-N, preparation method and application thereof
CN114044816B (en) * 2021-11-10 2023-06-16 大连海洋大学 Recombinant crassostrea gigas Jiao Kongsu protein rCgGSDME-N, preparation method and application thereof

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