CN106397554A - Cordyceps militaris lectin protein Lectin-ccm3, preparation method and application thereof - Google Patents
Cordyceps militaris lectin protein Lectin-ccm3, preparation method and application thereof Download PDFInfo
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- CN106397554A CN106397554A CN201610912122.7A CN201610912122A CN106397554A CN 106397554 A CN106397554 A CN 106397554A CN 201610912122 A CN201610912122 A CN 201610912122A CN 106397554 A CN106397554 A CN 106397554A
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Abstract
The invention relates to a cordyceps militaris lectin protein Lectin-ccm3, a preparation method and an application thereof. The amino acid sequence is shown as SEQ ID NO.1. The preparation method comprises the following steps: (1) linking cDNA sequence of the cordyceps militaris lectin protein into a vector, thereby obtaining a recombinant vector; (2) transferring the host cells from the recombinant vector, thereby obtaining a recombination strain; (3) culturing the recombination strain and inducing the expression of the recombinant cordyceps militaris lectin protein Lectin-ccm3; (4) purifying, dialyzing and separating, thereby obtaining the Lectin-ccm3. The Lectin-ccm3 is used for preparing an anti-microbial anti-cancer drug. According to the invention, the cordyceps militaris lectin protein Lectin-ccm3 is obtaining through in vitro recombination, the growth of staphylococcus aureus can be restrained, and the biological activity, such as multiplication of human cervical cancer cell HeLa, can be restrained, so that the cordyceps militaris lectin protein Lectin-ccm3 has an application potential at the anti-microbial anti-anticancer aspects.
Description
Technical field
The invention belongs to agglutinin field, particularly to a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 and its preparation
Methods and applications.
Background technology
Agglutinin (lectin) is that a class can optionally identify sugared and non-covalent Reversible binding therewith non-antibody protein
Matter.Because of its energy coagulation erythrocyte, therefore named agglutinin (Barcmdes, 1988).Nineteen ninety-five Peumans and Van Damme proposes
The new definition of phytohemagglutinin, that is, agglutinant protein contain at least one can with specificity monosaccharide or oligomeric (few) sugar reversibility knot
Close, on-catalytic domain (Peumans, 1995).
Cell agglutination is to be easiest to intuitively characterize the phenomenon that agglutinin is combined with cell.Agglutinin typically has two or two
Individual above sugared binding site, therefore can be with multiple specific cells in combination with so that free cell aggregation be agglomerating
And produce coagulation effect.Research shows, agglutinin not only can be with erythrocyte binding, can also be with antibacterial, lymphocyte, reproduction
Other cell such as cell combines.
Lectins from Mushrooms can the various erythrocyte of coagulation, tumor cell, lymphocyte etc..The cell agglutination of agglutinin
That the glycosyl of the subunit identification single-minded with the agglutinin of a cell surface of agglutinin molecule is combined, another subunit with another
Glycosyl on one cell combines, thus lead to (Liu Yanru, 2003) by the bridging action of agglutinin.
Agglutinin From Pleurotus Ostreatus to hepatoma H-22 (Wang, 2000), Ganoderma capenseD.A.Reid agglutinin to Ehrlich tumor and
M1 cell and hepatocarcinoma (HepG2) cell (Ngai, 2004), Fistulina hepatica agglutinin FHL are respectively provided with obvious suppression to HeLa cell etc.
Make and use (Lin Yuman, 2004).Phytohemagglutinin plays an important role in the defense reaction of plant, has antibacterium, funguses, elder brother
Worm, or even stop by the function (Peumans, 1995) of animal hunting.There is result of study display macro fungis agglutinin in funguses
In vivo may be similar to phytohemagglutinin, there is in the defense reaction of funguses important effect (Sun Hui, 2003).
The molecular biology research of agglutinin in Cordyceps militaris (L.) Link. be have not been reported, Cordyceps militaris (L.) Link. genome has been sequenced and has completed,
Annotation is carried out prokaryotic expression for agglutinin synthetic gene, builds the bacterial strain of agglutinin high efficient expression, specify Cordyceps militaris (L.) Link. agglutinin
Biological activity, be that the exploitation of agglutinin in Cordyceps militaris (L.) Link. lay the foundation.
Content of the invention
The technical problem to be solved is to provide a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 and its preparation
Methods and applications, this albumen can suppress the growth of staphylococcus aureuses, propagation of suppression human cervical carcinoma cell HeLa etc. raw
Thing activity, therefore, has application potential in antibacterial anticancer aspect.
The invention provides a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3, the aminoacid sequence such as SEQ of described albumen
Shown in IDNO.1.
Invention further provides a kind of gene of coding Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3, the sequence of described gene
Row are as shown in SEQID NO.2.
This albumen belongs to a kind of new mycoagglutinin albumen.Its cDNA sequence is carried out in GenBank BLAST ratio
Right, there is no to find a gene order similar to it, and by its aminoacid sequence NCBI irredundant protein sequence databank
(nr) carry out in BLAST compare find, this albumen with from Cordyceps brongniartii (Cordyceps brongniartii) agglutinin
The similarity of albumen reaches 84.79%, with the phase coming from beauveria bassiana (Beauveria bassiana) agglutinant protein
It is 85.19% like property, with the similarity coming from green muscardine funguss (Metarhizium brunneum) agglutinant protein be
51.49%.Illustrate that Lectin-ccm3 is a kind of new mycoagglutinin albumen, the gene of its coding is a new gene.
Invention further provides a kind of carrier of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene.
Preferably, described carrier is pET-32a.
Most preferably, EcoRI Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene being inserted on pET-32a limits
On property restriction enzyme site, obtain recombinant vector pET-Lectin-ccm3.
Invention further provides a kind of recombinant bacterial strain of the Lectin-ccm3 gene of agglutinant protein containing Cordyceps militaris (L.) Link..
Preferably, described recombinant bacterial strain is BL21-Lectin-ccm3.
A kind of preparation method of the Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 of the present invention, including：
(1) cDNA sequence of Cordyceps militaris (L.) Link. agglutinant protein is connected in carrier, obtains recombinant vector；
(2) by above-mentioned recombinant vector transformed host cell, obtain recombinant bacterial strain；
(3) above-mentioned recombinant bacterial strain, the expression of induction restructuring Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 are cultivated；
(4) purification, dialysis separate and obtain expressed Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3.
Host cell in described step (2) is e. coli bl21 cell.
A kind of application of the Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 of the present invention, described Cordyceps militaris (L.) Link. agglutinant protein
Lectin-ccm3 is used for preparing antibacterial cancer therapy drug.
Beneficial effect
The Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 that the present invention is obtained by vitro recombination can suppress golden yellow Fructus Vitis viniferae
The biological activitys such as the growth of coccus, the propagation of suppression human cervical carcinoma cell HeLa, therefore, have application in antibacterial anticancer aspect latent
Power.
Brief description
Fig. 1 is Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 aminoacid sequence of the present invention and other mycoagglutinin albumen
Amino acid alignment result；
Fig. 2 is pET-Lectin-ccm3 plasmid map of the present invention；
Fig. 3 is the SDS-PAGE analysis of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 expression of the present invention；Wherein, 1 is zero load
Body, 2 is not induce thalline, and 3 is bacterial cell disruption precipitation, and 4 is Lectin-ccm3 albumen, and M is molecular weight standard；
Fig. 4 is Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 suppression staphylococcus aureus growth experiment of the present invention；Its
In, Amp is positive control, antibacterial circle diameter 2.1cm, and 1 is empty carrier, and 2 is Lectin-ccm3 albumen, antibacterial circle diameter
1.2cm；
Fig. 5 is that Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 of the present invention suppresses HeLa cell proliferation experiment.
Specific embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than restriction the scope of the present invention.In addition, it is to be understood that after having read the content of present invention instruction, people in the art
Member can make various changes or modifications to the present invention, and these equivalent form of values equally fall within the application appended claims and limited
Scope.
Experimental technique in following embodiments, if no special instructions, is conventional method.Following is examination used in example
Test material, if no special instructions, be routine biochemistry Reagent Company and buy.Quantitative experiment in following examples, is respectively provided with three
Secondary repeat test, results averaged.
1. cell strain：BL21 competence, Top10 competence.
2. test kit, enzyme, biochemical reagents and instrument：
Test kit：Tiangeng plasmid extraction kit, Tiangeng protein quantification detection kit, the raw work PCR primer in Shanghai reclaims
Test kit, PCR primer synthesis and gene sequencing are completed by the raw work in Shanghai.
Enzyme：Takara routine restriction endonuclease, Takara DNA T4 ligase, Taq DNA polymerase
Instrument：Ai Bende micro sample adding appliance, match be silent fly your centrifuge of generation, Shanghai sky energy gel imaging instrument, Horizontal electrophoresis tank,
Vertial electrophorestic tank, Jiangyin Nuo Yuan constant-temperature shaking incubator, new sesame Ultrasonic Cell Disruptor, ABI PCR instrument, Bole's enzyme-linked immunosorbent assay instrument.
Biochemical reagents：
Traditional Chinese medicines glacial acetic acid, glycerol (glycerol), dehydrated alcohol, isopropanol, methanol, glycine, Tris, sodium hydroxide,
EDTA, sodium chloride, potassium chloride, 12 hypophosphite monohydrate sodium dihydrogens, potassium dihydrogen phosphate；
Acrylamide, N, N- methylene fork acrylamide, IPTG；Oxdine tryptone, yeast extract；Solarbio ammonia
Benzylpcnicillin, block that antibiotic；SDS, beta -mercaptoethanol, coomassie brilliant blue R_250, AP.
3. culture medium：Raw work LB culture medium.
Embodiment 1
Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene sequence information and homology analysis
Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 sequence of the present invention is 1299bp, and detailed sequence is shown in SEQ ID NO.2 institute
Show.Lectin-ccm3 aminoacid sequence is derived according to cDNA sequence, totally 432 amino acid residues, molecular weight 48.17kDa,
Theoretical isoelectric point, IP (pI) is 7.88, and unstability index is 32.26, and aliphatic coefficient is 79.00, and detailed sequence is shown in SEQ ID
Shown in NO.1.
By Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 aminoacid sequence blast program in Non-redundant
Carry out protein homology search in GenBank data base, it is found that it with from Cordyceps brongniartii (Cordyceps
Brongniartii) similarity of agglutinant protein reaches 84.79%, and comes from beauveria bassiana (Beauveria
Bassiana) similarity of agglutinant protein is 85.19%, and comes from green muscardine funguss (Metarhizium brunneum) coagulation
The similarity of fibroin is 51.49%.As shown in Figure 1.As can be seen here, Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 and funguses
There is higher homology in agglutinant protein family on protein level, be a kind of new mycoagglutinin albumen.
Embodiment 2
The amplification of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene and expression
CDNA is become for template reverse transcription with Cordyceps militaris (L.) Link. mRNA, then agglutinin gene is gone out for template amplification with above-mentioned cDNA
Lectin-ccm3, through EcoRI enzyme action, is connected to ligase (T4ligase) after purification and is carried with the pET32a digesting through EcoRI
On body, expression vector pET-Lectin-ccm3 is transformed into expression strain BL21.Picking monoclonal, LB fluid medium is cultivated, secondary
Day, strain is with 1：50 amplification culture 200mL, cultivate the IPTG adding 0.2mM to OD600=0.4-0.6,16 DEG C of inductions for 37 DEG C
Expression 12h.
Embodiment 3
The extraction of restructuring Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3
8000rpm, 4 DEG C of centrifugation 5min, collect the bacterial strain of abduction delivering, plus the broken liquid of 1mL are resuspended, laggard with liquid nitrogen flash freezer
Row grinds.12000rpm, 4 DEG C of centrifugation 15min, collect supernatant precipitation.The expression of SDS-PAGE testing goal albumen.Mesh
Albumen Lectin-ccm3 with soluble protein form exist, through biofilter degerming after -20 DEG C preservation.
Embodiment 4
Restructuring Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 suppression staphylococcus aureuses proliferation test
The golden yellow of incubated overnight after solid LB media melts, when temperature is down to 50 DEG C, is added with 1% inoculum concentration
Staphylococcuses bacterium solution, is down flat plate immediately after mixing, in surface Deca 2ul above-mentioned crude protein extracting solution after flat board solidification, with sky
Vector expression product as negative control, with ammonia benzyl as positive control.Flat board is placed in 37 DEG C cultivate 10 hours, observes knot
Really.
As shown in figure 4, purifying protein has inhibitory action to golden yellow glucose coccus.
Embodiment 5
Restructuring Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 suppression human cervical carcinoma cell HeLa proliferation test
Collect the HeLa cell growing to logarithmic (log) phase, adjustment concentration of cell suspension to 5*104Individual/mL, adds 96 orifice plates, often
Hole adds 100ul.5%CO2, 37 DEG C are incubated and are paved with bottom hole to cell monolayer, add above-mentioned crude protein extracting solution, every hole adds
100ul.5%CO2, 37 DEG C are incubated 24 hours, observation of cell growing way under inverted microscope.Every hole adds 20ul MTT solution
(5mg/ml, i.e. 0.5%MTT), continues culture 4h.Carefully suck in the hole culture fluid, every hole adds 150ul DMSO, puts on shaking table
Low-speed oscillation 10min, makes crystal fully dissolve.Measure the light absorption value in each hole at OD570nm in enzyme-linked immunosorbent assay instrument.
As shown in figure 5, when albumen crude extract concentration is 1.8mg/ml, suppression ratio is 17.89%.
Claims (10)
1. a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 it is characterised in that：The aminoacid sequence of described albumen such as SEQ ID
Shown in NO.1.
2. a kind of gene of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 as shown in claim 1 for the coding it is characterised in that：
The sequence of described gene is as shown in SEQ ID NO.2.
3. a kind of carrier containing Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene as claimed in claim 2.
4. the carrier of a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene according to claim 3, its feature exists
In：Described carrier is pET-32a.
5. the carrier of a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene according to claim 4, its feature exists
In：Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene is inserted on the EcoRI restriction enzyme site on pET-32a, obtains
To recombinant vector pET-Lectin-ccm3.
6. a kind of recombinant bacterial strain containing Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene as claimed in claim 2.
7. the recombinant bacterial strain of a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 gene according to claim 6, its feature
It is：Described recombinant bacterial strain is BL21-Lectin-ccm3.
8. a kind of preparation method of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3, including：
(1) cDNA sequence of Cordyceps militaris (L.) Link. agglutinant protein is connected into as, in the carrier of claim 3 or 4, obtained recombinant vector；
(2) by above-mentioned recombinant vector transformed host cell, obtain recombinant bacterial strain；
(3) above-mentioned recombinant bacterial strain, the expression of induction restructuring Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 are cultivated；
(4) purification, dialysis separate and obtain expressed Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3.
9. the preparation method of a kind of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 according to claim 8, its feature exists
In：Host cell in described step (2) is e. coli bl21 cell.
10. a kind of application of Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 as claimed in claim 1 it is characterised in that：Described
Cordyceps militaris (L.) Link. agglutinant protein Lectin-ccm3 is used for preparing antibacterial cancer therapy drug.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111875683A (en) * | 2020-07-31 | 2020-11-03 | 武汉轻工大学 | Cordyceps militaris agglutinin gene and preparation method thereof, recombinant expression vector, strain, and preparation method and application of cordyceps militaris agglutinin |
| WO2021224724A1 (en) | 2020-05-07 | 2021-11-11 | Unichem Laboratories Ltd | Anti-cancer proteins |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1560250A (en) * | 2004-03-04 | 2005-01-05 | 上海交通大学 | Coding sequence of Chinese yew agglutinant protein |
| CN102174524A (en) * | 2011-01-21 | 2011-09-07 | 四川大学 | Liparis nervosa lectin gene and protein as well as application thereof |
-
2016
- 2016-10-19 CN CN201610912122.7A patent/CN106397554A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1560250A (en) * | 2004-03-04 | 2005-01-05 | 上海交通大学 | Coding sequence of Chinese yew agglutinant protein |
| CN102174524A (en) * | 2011-01-21 | 2011-09-07 | 四川大学 | Liparis nervosa lectin gene and protein as well as application thereof |
Non-Patent Citations (4)
| Title |
|---|
| ZHENG,P.等: "Cordyceps militaris CM01 lectin family integral membrane protein, putative (CCM_01589), partial mRNA", 《GENBANK DATABASE》 * |
| ZHENG,P.等: "lectin family integral membrane protein, putative [Cordyceps militaris CM01]", 《GENBANK DATABASE》 * |
| 冯瑞娟 等: "半夏凝集素的分离纯化及其对Hela细胞生长的影响", 《浙江理工大学学报》 * |
| 张小团 等: "半夏蛋白的提取工艺及其凝集素的抑菌研究", 《西南师范大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021224724A1 (en) | 2020-05-07 | 2021-11-11 | Unichem Laboratories Ltd | Anti-cancer proteins |
| CN111875683A (en) * | 2020-07-31 | 2020-11-03 | 武汉轻工大学 | Cordyceps militaris agglutinin gene and preparation method thereof, recombinant expression vector, strain, and preparation method and application of cordyceps militaris agglutinin |
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Application publication date: 20170215 |