CN101698678B - Silkworm antibacterial peptide, cDNA sequence thereof and application thereof - Google Patents
Silkworm antibacterial peptide, cDNA sequence thereof and application thereof Download PDFInfo
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Abstract
The invention discloses silkworm antibacterial peptide BmcecE, a cDNA sequence thereof and application thereof. The amino acid sequence of the antibacterial peptide is expressed as SEQ ID NO:3, and the nucleotide sequence of coded cDNA is expressed as SEQ ID NO: 1. The silkworm antibacterial peptide has antibacterial activity for gram-positive bacterium (bacillus thuringiensis) and gram-negative bacterium (Escherichia coli, pseudomonas aeruginosa, prodigiosus septicemia or ralstonia solanacearum), and has broad antibacterial spectrum. The silkworm antibacterial peptide BmcecE can be used for medicament research and development for people, livestock and birds for preventing and treating diseases of people, livestock and birds caused by bacillus thuringiensis, Escherichia coli, pseudomonas aeruginosa, prodigiosus septicemia or ralstonia solanacearum infection; and the silkworm antibacterial peptide BmcecE also can be used for preparing biological product additives for preventing corruption degeneration of biological products caused by the bacillus thuringiensis, colon bacillus, pseudomonas aeruginosa, prodigiosus septicemia or ralstonia solanacearum.
Description
Technical field
The present invention relates to the domestic silkworm gene engineering research field, be specifically related to the research of cultivated silkworm antimicrobial peptide, relate in particular to a kind of new cultivated silkworm antimicrobial peptide and cDNA sequence and application.
Background technology
Insect has extremely strong adaptive faculty and defence capability; Wherein most important mechanism is to cause of disease material and harmful non-single-minded immune system that infects factor; Insect antimicrobial peptide (Antibacterial peptides); (Insect Antimicrobial Peptide AMP), mainly replys molecule as the non-single-minded immune response mechanism of insect body fluid to be called the insect antimicrobial peptide again; Demonstration has the killing action of broad-spectrum high efficacy to bacterium, fungi, to the restraining effect of virus, protozoon, improper cell (for example cancer cells).
The sterilization mechanism of the insect antimicrobial peptide mainly influence of the damage of the physical chemistry through cell membrane or cell membrane current potential or the breathing that suppresses bacteria cell wall reaches fungistatic effect; So be difficult for causing the microorganisms resistance; Have that molecular weight is little, thermally-stabilised, has a broad antifungal spectrum, non-immunogenicity, to advantage such as the higher animal normal cell is harmless; Possibly become and substitute one of antibiotic novel antibacterial material; So very active to the application of antibacterial peptide transgenic breeding for disease resistance and bio-pharmaceuticals (preparation) research and development both at home and abroad in recent years, up to now, isolation identification or found that kind more than 100 has the active insect antimicrobial peptide molecule of immune induction.
Cecropins family antibacterial peptide is one type of antibacterial peptide that research is more deep; Bibliographical information is all arranged from the insect to the vertebrates; This family is active stronger a kind of in the present known natural antibacterial peptide, from fruit bat (Drosophila melanogaster) and malarial mosquito Dipteras such as (Anopheles gambiae) and lepidopterous insect, has found the Cecropins gene family member that number does not wait respectively at present.
Cecropins family antibacterial peptide is about 4kD for the molecule amount; The alkaline heat-resisting small peptide of forming by 40 amino-acid residues (aa); Multiple homologue types such as A, B, C, D, E, F and G are arranged, and serve as typical case's representative with Cecropin A1-A3, B, the D that cherishs the guppy giant silkworm wherein.
Southwestern University had accomplished full order-checking and block diagram drawing (Xia etal. to the domestic silkworm gene group in 2004; 2004.Science; 306:1937-1940), for the research of cultivated silkworm antimicrobial peptide gene provide 9 * genome database and est database (silkworm knowledgebase:
Http:// silkworm.genomics. Org.cn/).
Summary of the invention
The object of the present invention is to provide a kind of new antibacterial peptide that derives from silkworm Cecropins family.
Another object of the present invention is to provide the cDNA sequence of above-mentioned antibacterial peptide.
Another object of the present invention is to provide the application of above-mentioned cultivated silkworm antimicrobial peptide.
Above-mentioned purpose of the present invention is achieved through following scheme:
The inventor is based on silkworm 9 * genome database and est database; Analyze and the DNAStar software analysis through SignalP online (http://www.cbs.dtu.dk/services/SignalP-2.0/); Found 1 new antibacterial peptide newcomer of silkworm Cecropins family, and with its called after BmcecE, the aminoacid sequence of this antibacterial peptide is shown in SEQ ID NO:3; Comprise 39 amino acid, the cDNA sequence of this antibacterial peptide is shown in SEQ ID NO:1.
Cultivated silkworm antimicrobial peptide BmcecA2 (mature peptide) molecule, cultivated silkworm antimicrobial peptide BmcecB1 (mature peptide) molecule and cultivated silkworm antimicrobial peptide Cecropin D (mature peptide) molecule, these three is the verified anti-microbial activity that has.
The present invention as experimental control, carries out anti-microbial activity research to the cultivated silkworm antimicrobial peptide BmcecE of above-mentioned acquisition with cultivated silkworm antimicrobial peptide BmcecA2 (mature peptide) molecule, cultivated silkworm antimicrobial peptide BmcecB1 (mature peptide) molecule and cultivated silkworm antimicrobial peptide Cecropin D (mature peptide) molecule.
Through discovering, cultivated silkworm antimicrobial peptide BmcecE of the present invention is to bacillus thuringiensis (Bacillusthuringiensis), intestinal bacteria K
12D
31(Escherichia coli K
12D
31), Pseudomonas aeruginosa (Pseudomonas aeruginosa), clever bacterium sepsis germ (Seiiatio maicesceis) and Ralstonia solanacearum (Ralstonia dolaanacearum) all have anti-microbial activity.
Cultivated silkworm antimicrobial peptide BmcecE is 1.25 μ M to the minimum inhibition concentration (MIC) of bacillus thuringiensis (Bacillus thuringiensis), to intestinal bacteria K
12D
31(Escherichia coli K
12D
31) minimum inhibition concentration (MIC) be 2.5 μ M; Minimum inhibition concentration (MIC) to Pseudomonas aeruginosa (Pseudomonas aeruginosa) is 2.5 μ M; Minimum inhibition concentration (MIC) to clever bacterium sepsis germ (Seiiatio maicesceis) is 2.5 μ M, is 2.5 μ M to the minimum inhibition concentration (MIC) of Ralstonia solanacearum (Ralstonia dolaanacearum).
Cultivated silkworm antimicrobial peptide BmcecE of the present invention can be used for the medicament research and development of people, animal, fowl, and prevention and treatment are infected by bacillus thuringiensis, intestinal bacteria, Pseudomonas aeruginosa, clever bacterium sepsis germ or Ralstonia solanacearum and caused causing a disease of people, animal, fowl; Can also be used for the preparation of biologics additive, the biologics that prevents to cause putrid and deteriorated by bacillus thuringiensis, intestinal bacteria, Pseudomonas aeruginosa, clever bacterium sepsis germ or Ralstonia solanacearum.
Compared with prior art, the present invention has following beneficial effect:
1. the present invention obtains a new cultivated silkworm antimicrobial peptide, has enriched existing cultivated silkworm antimicrobial peptide kind, for the research of cultivated silkworm antimicrobial peptide provides new kind;
2. cultivated silkworm antimicrobial peptide of the present invention is to gram-positive microorganism (bacillus thuringiensis); And Gram-negative bacteria (intestinal bacteria, Pseudomonas aeruginosa, clever bacterium sepsis germ or Ralstonia solanacearum) all has anti-microbial activity; This antibacterial peptide has a broad antifungal spectrum is described, is had more wide application prospect;
3. the medicament research and development that cultivated silkworm antimicrobial peptide of the present invention can be used for being correlated with and the preparation of biological products additive adopt gene engineering method to produce this antimicrobial peptide protein matter, have higher commercial value.
Description of drawings
Fig. 1 is the electrophorogram of the Sephadex G-10 chromatography desalting and purifying among the embodiment 2;
Wherein, M is Marker, and 1 is bacterial lysate, and 2 is BmcecA2, the 3rd, and BmcecE, 4 is BmcecB1,5 is Cecropin D;
Fig. 2 is the single protein electrophorese figure among the embodiment 2;
Wherein, M is Marker, and 1 is BmcecA2, the 2nd, and BmcecE, 3 is BmcecB1,4 is Cecropin D.
Embodiment
Below in conjunction with specific embodiment the present invention is done description further, but specific embodiment is not done any qualification to the present invention.
The acquisition of embodiment 1 silkworm novel polypeptide BmcecE
The inventor is based on silkworm 9 * genome database and est database; Analyze and the DNAStar software analysis through SignalP online (http://www.cbs.dtu.dk/services/SignalP-2.0/); Found 1 new silkworm polypeptide, and with its called after BmcecE, this amino acid sequence of polypeptide is shown in SEQ IDNO:3; Comprise 39 amino acid, the cDNA sequence of this polypeptide is shown in SEQ ID NO:1.
The structure of embodiment 2 recombinant expression vector plasmids, transformant bacterial strain and protein expression
Cultivated silkworm antimicrobial peptide BmcecA2 (mature peptide) molecule, cultivated silkworm antimicrobial peptide BmcecB1 (mature peptide) molecule and cultivated silkworm antimicrobial peptide Cecropin D (mature peptide) molecule; These three is the verified anti-microbial activity that has; Wherein, the nucleotide sequence of cultivated silkworm antimicrobial peptide BmcecA2 gene, its GenBank accession number is NM_001043997; The nucleotide sequence of cultivated silkworm antimicrobial peptide BmcecB1 gene; Its GenBank accession number is 1703262B/AAC60501, the nucleotide sequence of cultivated silkworm antimicrobial peptide Cecropin D gene, and its GenBank accession number is BAA31507.
Cultivated silkworm antimicrobial peptide BmcecA2, cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D and embodiment 1 gained silkworm polypeptide BmcecE, the aminoacid sequence comparison of these four mature peptides is as follows:
BmcecA2 RWKLFKKIEKVGRNVRDGLIKAGPAIAVIGQAKSL 35aa
BmcecB1 RWKIFKKIEKMGRNIRDGIVKAGPAIEVLGSAKAIGK 37aa
CecropinD GNFFKDLEKMGQRVRDAVISAAPAVDTLAKAKALGQG 37aa
BmcecE RWKIFKKIEKVGQNIRDGIIKAGPAVAVVGQAATIAHGK 39aa
* * ** * *** *
In the above-mentioned contrast, on behalf of N end basic aminoacids, the A-G-P amino acid that can form the β break over region and C, asterisk (*) hold amidable amino acid sites respectively.
From above-mentioned contrast, can find out; Embodiment 1 gained cultivated silkworm antimicrobial peptide BmcecE has higher similarity with existing antibacterial peptide on aminoacid sequence; Especially some maybe with the active relevant site of antibacterial on; Also be to keep identical, so the inventor predict that embodiment 1 gained silkworm polypeptide BmcecE should also have certain anti-microbial activity.
As experimental control, the silkworm polypeptide BmcecE that the foregoing description 1 is obtained carries out anti-microbial activity research to present embodiment with cultivated silkworm antimicrobial peptide BmcecA2 (mature peptide) molecule, cultivated silkworm antimicrobial peptide BmcecB1 (mature peptide) molecule and cultivated silkworm antimicrobial peptide Cecropin D (mature peptide) molecule.
1, the acquisition of design of primers and cDNA sequence
According to cultivated silkworm antimicrobial peptide BmcecA2 gene, cultivated silkworm antimicrobial peptide BmcecB1 gene, cultivated silkworm antimicrobial peptide Cecropin D gene and silkworm polypeptide BmcecE gene, design corresponding amplimer respectively, deliver biotech firm and synthesize.
As template, adopt above-mentioned amplimer with silkworm pupa fatty body RNA respectively, through conventional RT-PCR amplification operation, amplification obtains the cDNA sequence of above-mentioned four gene encoding mature peptides.
The primer and the cDNA of present embodiment are as follows:
(1) cultivated silkworm antimicrobial peptide BmcecA2 gene
Amplimer:
BmcecA2-S, its nucleotide sequence is shown in SEQ ID NO:4;
BmcecA2-A, its nucleotide sequence is shown in SEQ ID NO:5.
The cDNA length of amplification gained cultivated silkworm antimicrobial peptide BmcecA2 gene encoding mature peptide is 131bp.
(2) cultivated silkworm antimicrobial peptide BmcecB1 gene
Amplimer:
BmcecB1-S, its nucleotide sequence is shown in SEQ ID NO:6;
BmcecB1-A, its nucleotide sequence is shown in SEQ ID NO:7.
The cDNA length of amplification gained cultivated silkworm antimicrobial peptide BmcecB1 gene encoding mature peptide is 130bp.
(3) cultivated silkworm antimicrobial peptide Cecropin D gene
Amplimer:
Cecropin D-S, its nucleotide sequence is shown in SEQ ID NO:8;
Cecropin D-A, its nucleotide sequence is shown in SEQ ID NO:9.
The cDNA length of amplification gained cultivated silkworm antimicrobial peptide Cecropin D gene encoding mature peptide is 117bp.
(4) silkworm polypeptide BmcecE gene
Amplimer:
BmcecE-S, its nucleotide sequence is shown in SEQ ID NO:10;
BmcecE-A, its nucleotide sequence is shown in SEQ ID NO:11.
The cDNA length of amplification gained silkworm polypeptide BmcecE gene encoding mature peptide is 130bp.
In above-mentioned each primer, CCATGG is a Nco I restriction enzyme site, and CTCGAG is an Xho I restriction enzyme site, and TCA is a terminator.
2, the structure of recombinant expression vector plasmid, the acquisition of transformant bacterial strain
Through two restriction endonuclease sites Nco I and the Xho I that adds on the above-mentioned PCR primer; The cDNA fragment of four genes of above-mentioned amplification gained is connected respectively in expression vector pET-32a (+) plasmid, makes up and obtain the recombinant expression vector plasmid: plasmid pET-32a (+)-BmcecA2, pET-32a (+)-BmcecB1, pET-32a (+)-Cecropin D and pET-32a (+)-BmcecE.
With above-mentioned recombinant expression vector plasmid difference Transformed E .coli Rosetta
TM(DE3) behind the recipient bacterium, identify and the order-checking evaluation, obtain to contain transformant bacterial strain pET-32a (+)-BmcecA2/E.coli Rosetta of recombinant expression vector plasmid through conventional PCR
TM(DE3), pET-32a (+)-BmcecB1/E.coliRosetta
TM(DE3), pET-32a (+)-Cecropin D/E.coli Rosetta
TM(DE3) and pET-32a (+)-BmcecE/E.coli Rosetta
TM(DE3).
3, protein expression
The above-mentioned transformant bacterial strain that respectively contains the recombinant expression vector plasmid is carried out target protein BmcecA2 (B1, E, D, abduction delivering cultivation (starter bacteria liquid OD D1) respectively
6000.8; IPTG concentration 0.6mmol/L induces 4h); Through N,O-Diacetylmuramidase cracking, ultrasonic disruption, centrifugal collection contains the warm proteic supernatant of cultivated silkworm antimicrobial peptide BmcecA2 (cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D or cultivated silkworm antimicrobial peptide BmcecE) of mainly expressing with soluble form to culture.
In the present embodiment; The cDNA sequence of cultivated silkworm antimicrobial peptide BmcecA2 (cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D or cultivated silkworm antimicrobial peptide BmcecE) is inserted between the Nco I and Xho II of expression vector pET-32a (+); Make itself and Trx (Trx) gene fusion expression; In the warm albumen of its expression, there is the Trx-HisTag-Thrombin-STag-EK cleavage site; So; To containing the warm albumen supernatant of cultivated silkworm antimicrobial peptide BmcecA2 (cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D or cultivated silkworm antimicrobial peptide BmcecE); At first through the HisTag in the warm albumen (histidine-tagged) and the specific combination of Ni-NTA affinity column and the separation and purification of Sephadex G-10 chromatography desalting treatment, obtain the warm proteic purification of samples of cultivated silkworm antimicrobial peptide BmcecA2 (cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide CecropinD or cultivated silkworm antimicrobial peptide BmcecE), the electrophorogram of Sephadex G-10 chromatography desalting and purifying is as shown in Figure 1.
By enteropeptidase (EK) the warm proteic purification of samples of above-mentioned cultivated silkworm antimicrobial peptide BmcecA2 (cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D or cultivated silkworm antimicrobial peptide BmcecE) is cut at the EK cleavage site; Then cleaved products is carried out 2 molecule ultrafiltration respectively; Promptly cross post and remove the above Trx (Trx) of 10.0kD with the ultrafiltration pipe of molecular weight cut-off 10.0kD; Collect bottom solution; Use molecular weight cut-off to cross the single protein that post obtains the cultivated silkworm antimicrobial peptide BmcecA2 (cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D or cultivated silkworm antimicrobial peptide BmcecE) that molecular weight is about 4.0kD again as the ultrafiltration pipe of 3kD, as shown in Figure 2.
Can find out from Fig. 1 and Fig. 2, the protein of silkworm BmcecE genetic expression, its size is identical with cultivated silkworm antimicrobial peptide BmcecA2, cultivated silkworm antimicrobial peptide BmcecB1 and cultivated silkworm antimicrobial peptide Cecropin D.
Present embodiment is to the single protein of the cultivated silkworm antimicrobial peptide BmcecA2 (cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D or cultivated silkworm antimicrobial peptide BmcecE) of acquisition among the embodiment 2; Adopt conventional agar plate hole diffusion process (dull and stereotyped inhibition zone method) and conventional minimum inhibition concentration method (MIC), with following 5 kinds of Gram-negative (G
-), positive (G
+) bacterium is as being tried bacterium, carries out the test of anti-microbial activity, the result is shown in table 1 and table 2.
Bacillus thuringiensis (Bacillus thuringiensis) (G
+)
Intestinal bacteria K
12D
31(Escherichia coli K
12D
31) (G
-)
Pseudomonas aeruginosa (Pseudomonas aeruginosa) (G
-)
Spirit bacterium sepsis germ (Seiiatio maicesceis) (G
-)
Ralstonia solanacearum (Ralstonia dolaanacearum) (G
-)
Table 1 antibacterial circle diameter (mm)
| Supply the examination bacterium | BmcecB1 | Cecropin D | BmcecA2 | BmcecE | CK |
| Bacillus thuringiensis | 5.167±0.153 | 4.000±0.265 | 3.033±0.208 | 6.033±0.153 | - |
| Intestinal bacteria K 12D 31 | 8.600±0.200 | 5.100±0.173 | 3.967±0.208 | 4.033±0.252 | - |
| Spirit bacterium sepsis germ | 5.267±0.153 | 3.133±0.208 | - | 3.067±0.153 | - |
| Pseudomonas aeruginosa | 6.933±0.153 | 4.133±0.208 | 3.100±0.200 | 4.133±0.208 | - |
| Ralstonia solanacearum | 6.067±0.252 | 4.933±0.231 | 4.167±0.252 | 3.000±0.200 | - |
In the above-mentioned table 1, sample concentration is 5 μ mol/L, and CK is contrast (sterilization distilled water), and-expression does not have anti-microbial activity, and numeric representation is three multiple mean number ± standard errors in the table.
Table 2 minimum inhibition concentration (MIC) (μ M)
| Supply the examination bacterium | BmcecB1 | Cecropin D | BmcecA2 | BmcecE |
| Bacillus thuringiensis | 1.25 | 1.25 | 2.5 | 1.25 |
| Intestinal bacteria K 12D 31 | 0.625 | 1.25 | 2.5 | 2.5 |
| Spirit bacterium sepsis germ | 0.625 | 2.5 | - | 2.5 |
| Pseudomonas aeruginosa | 0.625 | 1.25 | 2.5 | 2.5 |
| Ralstonia solanacearum | 1.25 | 1.25 | 2.5 | 2.5 |
In the above-mentioned table 2 ,-expression is not carried out MIC and is measured.
Can find out from table 1 and table 2: surveying the result with giving birth to of 10 μ L same concentrations (5 μ mol/L) antibacterial peptide, silkworm antibacterial peptide BmcecA2, cultivated silkworm antimicrobial peptide BmcecB1, cultivated silkworm antimicrobial peptide Cecropin D4 and silkworm polypeptide BmcecE are tried bacterium to 5 kinds and are all shown tangible anti-microbial activity.
In conjunction with the result of study of embodiment 2 and embodiment 3, the silkworm polypeptide BmcecE of embodiment 1 belongs to silkworm Cecropins family, is newfound a kind of antibacterial peptide.
Cultivated silkworm antimicrobial peptide and cDNA sequence thereof and application sequence table
SEQUENCE LISTING
< 110>Ji'nan University, Southwestern University, Agricultural University Of South China
< 120>cultivated silkworm antimicrobial peptide and cDNA sequence and application
<130>
<160>11
<170>PatentIn version 3.5
<210>1
<211>120
<212>DNA
< 213>silkworm (Bombyx mori)
<400>1
agatggaaga ttttcaagaa aatcgaaaag gtgggtcaga acattcgtga tgggataatc 60
aaggctggac cagctgtcgc ggtggtaggg caggcggcga ccatcgctca cgggaaataa 120
<210>2
<211>120
<212>DNA
< 213>silkworm (Bombyx mori)
<220>
<221>CDS
<222>(1)..(117)
<400>2
aga tgg aag att ttc aag aaa atc gaa aag gtg ggt cag aac att cgt 48
Arg Trp Lys Ile Phe Lys Lys Ile Glu Lys Val Gly Gln Asn Ile Arg
1 5 10 15
gat ggg ata atc aag gct gga cca gct gtc gcg gtg gta ggg cag gcg 96
Asp Gly Ile Ile Lys Ala Gly Pro Ala Val Ala Val Val Gly Gln Ala
20 25 30
gcg acc atc gct cac ggg aaa taa 120
Ala Thr Ile Ala His Gly Lys
35
<210>3
<211>39
<212>PRT
< 213>silkworm (Bombyx mori)
<400>3
Arg Trp Lys Ile Phe Lys Lys Ile Glu Lys Val Gly Gln Asn Ile Arg
1 5 10 15
Asp Gly Ile Ile Lys Ala Gly Pro Ala Val Ala Val Val Gly Gln Ala
20 25 30
Ala Thr Ile Ala His Gly Lys
35
<210>4
<211>30
<212>DNA
< 213>artificial sequence
<400>4
catgccatgg ttaggtggaa actcttcaag 30
Cultivated silkworm antimicrobial peptide and cDNA sequence thereof and application sequence table
<210>5
<211>36
<212>DNA
< 213>artificial sequence
<400>5
gatatctcga gtcataagga tttcgcttgc cctatg 36
<210>6
<211>30
<212>DNA
< 213>artificial sequence
<400>6
catgccatgg ttaggtggaa gatcttcaag 30
<210>7
<211>35
<212>DNA
< 213>artificial sequence
<400>7
gaccctcgag tcagatagct ttagccgaac caagg 35
<210>8
<211>28
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< 213>artificial sequence
<400>8
cagtccatgg gcaacttctt caaggatc 28
<210>9
<211>35
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< 213>artificial sequence
<400>9
ccgctcgagt cattgtccga gagcttttgc ttttg 35
<210>10
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catgccatgg ttagatggaa gattttcaag 30
<210>11
<211>30
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< 213>artificial sequence
<400>11
atatctcgag tcagatggtc gccgcctgcc 30
Claims (4)
1. cultivated silkworm antimicrobial peptide BmcecE, its aminoacid sequence is shown in SEQ ID NO:3.
2. the cDNA of coding claim 1 said cultivated silkworm antimicrobial peptide BmcecE, its nucleotide sequence is shown in SEQ ID NO:1.
3. the application of the said cultivated silkworm antimicrobial peptide BmcecE of claim 1 in preparation treatment or prevention bacillus thuringiensis, intestinal bacteria, Pseudomonas aeruginosa, clever bacterium sepsis germ or Ralstonia solanacearum medicine.
4. the application of the said cultivated silkworm antimicrobial peptide BmcecE of claim 1 in the anti-bacillus thuringiensis of preparation, intestinal bacteria, Pseudomonas aeruginosa, clever bacterium sepsis germ or Ralstonia solanacearum biologics additive.
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| CN101698678B true CN101698678B (en) | 2012-01-11 |
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ID=42147142
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1158382C (en) * | 2001-02-28 | 2004-07-21 | 华南农业大学 | Preparation and application of antibiotic peptide pichia yeast |
| CN1587384A (en) * | 2004-08-27 | 2005-03-02 | 广州维观生物科技有限公司 | Antibiotic peptide gene converted viscons bacillus engineering bacterium and its preparing method and use |
-
2009
- 2009-11-03 CN CN2009101936200A patent/CN101698678B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1158382C (en) * | 2001-02-28 | 2004-07-21 | 华南农业大学 | Preparation and application of antibiotic peptide pichia yeast |
| CN1587384A (en) * | 2004-08-27 | 2005-03-02 | 广州维观生物科技有限公司 | Antibiotic peptide gene converted viscons bacillus engineering bacterium and its preparing method and use |
Non-Patent Citations (2)
| Title |
|---|
| Sun-Mee Hong et al..Structure and Expression Analysis of the Cecropin-E Gene from the Silkworm,Bombyx mori.《Biosci.Biotechnol.biochem.》.2008,第72卷(第8期), * |
| Tingcai Cheng et al..Structures, regulatory regions, and inductive expression patterns of antimicrobial peptide genes in the silkworm Bombyx mori.《Genomics》.2006,第87卷 * |
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