CN113831399B - Application of short peptide LBD-Q in preparation of antiviral infection product, feed and medicine - Google Patents

Application of short peptide LBD-Q in preparation of antiviral infection product, feed and medicine Download PDF

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CN113831399B
CN113831399B CN202110616966.8A CN202110616966A CN113831399B CN 113831399 B CN113831399 B CN 113831399B CN 202110616966 A CN202110616966 A CN 202110616966A CN 113831399 B CN113831399 B CN 113831399B
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CN113831399A (en
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张鹤千
余祥勇
毛勇
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Zhuhai Campus Of Beijing Normal University
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
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    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
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Abstract

The invention relates to the technical field of biology, in particular to application of short peptide LBD-Q in preparation of an anti-virus infection product, feed and a medicament. The inventor of the invention discovers for the first time through experiments that the short peptide LBD-Q containing the amino acid sequence shown as SEQ ID No.2 has an antiviral effect, can remarkably improve the antiviral effect of a product when being applied to an antiviral infection product, and particularly has strong clearing capacity on WSSV viruses which are easy to infect penaeus japonicus.

Description

Application of short peptide LBD-Q in preparation of antiviral infection product, feed and medicine
Technical Field
The invention relates to the technical field of biology, in particular to application of short peptide LBD-Q in preparation of an anti-virus infection product, feed and a medicament.
Background
Invertebrates mainly or completely depend on the innate immune system to resist infection or invasion of pathogenic organisms, wherein antibacterial peptides (AMPs) play an extremely important role, and many researches find that the prawn AMPs have strong antibacterial and antiviral activities and are concerned about due to potential application prospects and the function of killing bacteria or viruses in the aquaculture industry. The crustacean antibacterial peptide is mainly divided into chitin (Crustins), prawnin (Penaeidins) and anti-lipopolysaccharide factor (ALFs), and has certain antibacterial or antiviral activity. ALFs are one of the important components in antibacterial peptides of prawns, and are important and key components for host defense against bacteria, fungi and viruses.
Antimicrobial peptides are attractive candidates and feed additives due to their selectivity, speed of action and resistance to bacterial development. Natural and synthetic antimicrobial peptides are one of the main options for overcoming drug resistance. Due to the increasing resistance of antibiotics, new treatments for infections are urgently needed. The antibacterial peptide provides an opportunity for the antibacterial peptide, and in recent years, identification, optimization and modification of the antibacterial peptide arouse great interest of researchers. To date, more and more antimicrobial peptides from different species have been identified and uploaded into the database ADB of antimicrobial peptides. In general, the basic structure of the current antibacterial peptides can be divided into the following basic structure types, i.e., alpha helix, beta sheet, loop structure and stretch structure. Based on these four structures, a relatively wide variety of antimicrobial peptides are constructed, and understanding the structure helps to better explain the activity of the antimicrobial peptides.
However, the activity of many natural antibacterial peptides identified and separated at present is not ideal or has certain toxicity to eukaryotic cells, and the application and development of the antibacterial peptides at present mainly aim at resisting bacteria, so that the attention on the antibacterial capability of the antibacterial peptides is not enough, the research and development investment is low, and antibacterial peptide products with high antiviral capability are not found.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide application of short peptide LBD-Q in preparing products for resisting virus infection, and provide feed and medicaments containing the short peptide LBD-Q, so as to make up for the blank of the prior art which lacks antiviral products containing antibacterial peptide.
In order to solve the technical problems and realize the technical problems, the invention adopts the following technical scheme:
in a first aspect, the invention provides an application of a short peptide LBD-Q in preparation of an anti-viral infection product, wherein the amino acid sequence of the short peptide LBD-Q comprises a sequence shown as SEQ ID No.2, the sequence shown as SEQ ID No.2 is an artificial sequence obtained by designing and modifying an LBD short peptide of MjALF-D, and the amino acid sequence of the LBD short peptide of MjALF-D is a sequence shown as SEQ ID No. 1.
In an alternative embodiment, the two cysteines in the sequence shown in SEQ ID No.2 are linked by a disulfide bond.
In alternative embodiments, the sequence set forth in SEQ ID No.2 has protective modifications at both ends; preferably, the protective modification comprises a chemical modification.
In alternative embodiments, the amino terminus of the sequence depicted in SEQ ID No.2 is modified by acetylation.
In an alternative embodiment, the carboxy terminus of the sequence shown in SEQ ID No.2 is amidated.
In alternative embodiments, the virus comprises a WSSV virus.
In an alternative embodiment, the anti-viral infection product comprises a medicament or a feed.
In a second aspect, the present invention provides a feed comprising a main material, an auxiliary material and an additive, wherein the additive comprises the short peptide LBD-Q according to any one of the preceding embodiments.
Preferably, the main material comprises at least one of seeds, bran or cake.
Preferably, the adjuvant comprises at least one of a binder, a mildewcide, or an antioxidant.
Preferably, the additive further comprises at least one of mineral element additives, animal-derived protein additives, vitamin additives, amino acid additives, allicin additives or growth factor additives.
Preferably, the short peptide LBD-Q is coupled to an outer membrane protein.
Preferably, the outer membrane protein is derived from salmonella.
Preferably, the feed comprises a shrimp farming feed.
Preferably, the shrimp comprises marsupenaeus japonicus.
In a third aspect, the present invention provides a medicament comprising the short peptide LBD-Q according to any of the preceding embodiments and a pharmaceutically acceptable excipient.
Preferably, the short peptide LBD-Q is coupled to an outer membrane protein.
Preferably, the outer membrane protein is derived from salmonella.
Preferably, the medicament is in a dosage form comprising a suspension, a granule, a spray, an emulsion or a medicated bath.
In an alternative embodiment, the adjuvant for the drug comprises a cosolvent, an antioxidant, a pH adjuster, a solvent, and a penetrant.
The inventor finds out through experiments for the first time that the short peptide LBD-Q provided by the invention has an antiviral effect, can obviously improve the antiviral effect of a product when being applied to an antiviral infection product, and particularly has strong clearing capacity on WSSV viruses which are easy to infect penaeus japonicus.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a reversed-phase high performance liquid chromatogram of antibacterial short peptide LBD-Q in example 2;
FIG. 2 is a report of mass spectrometry analysis of short antibacterial peptide LBD-Q in example 2;
FIG. 3 shows the results of mass spectrometric detection of synthesized TaqMan probe-WSSV 32706;
FIG. 4 is a graph showing the cumulative mortality of Penaeus japonicus after the treatment of co-incubation of LBD-Q short peptide and WSSV in Experimental example 1;
FIG. 5 shows the results of detecting the number of copies of viruses in Experimental example 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
In addition, unless otherwise specified, the amino acid sequence shown in the present invention has a carboxyl terminus at the left end and an amino terminus at the right end.
In a first aspect, the invention provides an application of a short peptide LBD-Q in preparation of an anti-viral infection product, wherein the amino acid sequence of the short peptide LBD-Q comprises a sequence shown as SEQ ID No.2, the sequence shown as SEQ ID No.2 is an artificial sequence obtained by designing and modifying an LBD short peptide of MjALF-D, and the amino acid sequence of the LBD short peptide of MjALF-D is a sequence shown as SEQ ID No. 1. ALF is an effector molecule with broad-spectrum antibacterial activity in crustaceans, an anti-lipopolysaccharide factor found in prawns is named MjALF, a plurality of MjALFs are reported at present, including MjALF-A, MjALF-B, MjALF-D2, MjALF-E and the like, the existing reports mostly carry out research on the antibacterial and antiviral activities of the plurality of naturally-occurring MjALFs, and the MjALF is rarely artificially designed and improved in purpose.
In an alternative embodiment, the two cysteines in the sequence shown in SEQ ID No.2 are linked by a disulfide bond.
In alternative embodiments, the sequence set forth in SEQ ID No.2 has protective modifications at both ends; preferably, the protective modification comprises a chemical modification.
In alternative embodiments, the amino terminus of the sequence depicted in SEQ ID No.2 is modified by acetylation.
In an alternative embodiment, the carboxy terminus of the sequence shown in SEQ ID No.2 is amidated.
The disulfide bond connection and the protective chemical modification can obviously improve the stability of the short peptide against the influence of external factors, so that the short peptide is not easily degraded when being used as a feed or a drug additive, ensures the stable absorption of the short peptide and plays an antiviral role.
In alternative embodiments, the virus comprises a WSSV virus.
The WSSV virus is a non-inclusion body virus, because different scholars have different research methods and focus points, the calling methods for the virus are different, but the achieved basic consensus is that the virus is the pathogen causing the fulminant epidemic disease of the prawns, at present, a plurality of early rapid detection technologies are preliminarily established, including the application of a nested PCR detection technology, but as for prevention and treatment measures, the risk of the prawn infection can be reduced as much as possible only by strengthening the measures of desilting and disinfection, environmental management, pool inspection and the like, and effective medicine can be avoided.
In an alternative embodiment, the anti-viral infection product comprises a medicament or a feed.
In a second aspect, the present invention provides a feed comprising a main material, an auxiliary material and an additive, wherein the additive comprises the short peptide LBD-Q according to any one of the preceding embodiments.
Preferably, the main material comprises at least one of seeds, bran or cake.
Preferably, the adjuvant comprises at least one of a binder, a mildewcide, or an antioxidant.
The adhesive can bind some of the medicament powders which are not inherently sticky or less sticky. The binder may be, for example, but not limited to, alcohol, methyl cellulose, polyvinylpyrrolidone K30, polyvinylpyrrolidone K15, or starch slurry.
Mildewcides are a class of additives that inhibit the growth of and kill mold, including, but not limited to, ammonium persulfate, calcium phosphate, zinc ion, silver ion, or copper ion compounds.
The antioxidant can protect the medicinal components from oxidative damage and deterioration. The antioxidant may be, for example, but not limited to, sodium sulfite, sodium thiosulfate, or EDTA-2 Na.
Preferably, the additive further comprises at least one of mineral element additives, animal-derived protein additives, vitamin additives, amino acid additives, allicin additives or growth factor additives.
The feed is mainly used for feeding prawns, and antiviral ability of the prawns in the feeding process is improved by adding antiviral short peptide LBD-Q on the basis of common raw materials of the feed for the prawns.
Preferably, the short peptide LBD-Q is coupled to an outer membrane protein.
Preferably, the outer membrane protein is derived from salmonella.
The outer membrane protein of salmonella is a target protein of a plurality of bacteriophage fiber proteins, and the short peptide LBD-Q is coupled with the outer membrane protein, so that the short peptide LBD-Q can quickly target WSSV virus, and the effective rate of the short peptide LBD-Q before degradation is obviously improved.
Preferably, the feed comprises a shrimp farming feed.
Preferably, the shrimp comprises marsupenaeus japonicus.
The marsupenaeus japonicus is the most important breed of the penaeus japonicus, the penaeus japonicus is cultivated in Japan until the sale price is about 25 g, and the live penaeus japonicus is mainly sold. The culture of the Chinese Fujian, Guangdong and other Chinese south coastal areas has also started in recent years. The weight of the cultivated fish can reach 20-25 g in 180 days. The length of the spawning colony is 12-20 cm, the weight of the spawning colony is 20-95 g, the shrimp shell is thick, the shrimp shell is resistant to exposure, the shrimp shell is suitable for live transportation and marketing, the profit is high, and the nutritional value of the shrimp shell is close to that of other main shrimps.
The disease prevention is a key step in the breeding of the marsupenaeus japonicus, the traditional method is that the quick lime water is used for splashing 1 time in the whole pond every 10 to 15 days in the middle and later period of the breeding of the marsupenaeus japonicus, the physicochemical factor of the pond water is improved, the disease is prevented, and the consumption of the quick lime is 15ppm to 20 ppm. The terramycin is added into the bait to prepare the medicinal bait which can prevent bacterial diseases, and the addition amount is generally 1 per mill of the bait. In recent years, viral leukoderma is also generated in the process of culturing the marsupenaeus japonicus, an effective treatment means for viral diseases is lacked, and the generation can be slowed only by a prevention means, such as feeding high-quality bait to enhance the physique of the prawns, adding immune enhancing substances such as vitamin C, vitamin E, ginsenoside and the like into the bait to improve the immunity of the prawns. The pathogen of the shrimp white spot syndrome virus is a non-inclusion body virus, and the short peptide LBD-Q provided by the invention has a remarkable inhibiting effect on shrimp viruses through verification.
In a third aspect, the present invention provides a medicament comprising the short peptide LBD-Q according to any of the preceding embodiments and a pharmaceutically acceptable excipient.
Preferably, the short peptide LBD-Q is coupled to an outer membrane protein.
Preferably, the outer membrane protein is derived from salmonella.
Preferably, the medicament is in a dosage form comprising a suspension, a granule, a spray, an emulsion or a medicated bath.
In an alternative embodiment, the adjuvant for the drug comprises a cosolvent, an antioxidant, a pH adjuster, a solvent, and a penetrant.
The cosolvent and the difficultly soluble medicine form soluble intermolecular complex, association or double salt and the like so as to increase the solubility of the medicine in the solvent. The cosolvent of the granule may be, for example, but not limited to, arginine, histidine, methionine, meglumine, betacyclodextrin, tween 80, polyethylene glycol 400, or polyethylene glycol 600.
The pH regulator is used for maintaining or changing the pH value of the medicine. The pH adjuster may be, for example, but is not limited to, sodium hydroxide.
The solvent has stable physicochemical properties, can increase the solubility of the drug, can be physiologically compatible with body tissues, and can be quickly absorbed.
Penetrants can help some functional components penetrate into the stratum corneum of the skin to exert effects. The osmotic agent may be, for example, but is not limited to, ethanol or laurocapram.
Some embodiments of the invention are described in detail below with reference to the accompanying drawings. The embodiments described below and the features of the embodiments can be combined with each other without conflict.
EXAMPLE 1 engineering of antimicrobial peptides LBD short peptides based on MjALF-D
According to The Analysis of The physicochemical properties and The structural information of The LBD short Peptide of MjALF-D, The Analysis Software uses The ProtParam online Analysis Software (http:// www.expasy.org/tools/ProtParam) in The Exterprotein Analysis Software (ExPASY) website and The antibacterial Peptide Database (APD, http:// APs. unmac. edu/AP/main. php) online Analysis Software to obtain The relevant parameters of The polypeptide. Analysis of the arrangement of the hydrophilic and hydrophobic amino acids of the short peptides and helical wheel projection were performed using the online analysis software Heliquest analysis (http:// helix. ipmc. cnrs. fr/cgi-bin/ComputParams. py) (Gautier et al, 2008). The sequence of the MjALF-D LBD short peptide (LBD-ALF) is compared in an APD antibacterial peptide system, amino acid substitution is carried out according to the comparison result and all obtained relevant parameters, so that the activity of the short peptide is improved, a new LBD short peptide is reconstructed and designed, and the physicochemical property of the new designed short peptide LBD-Q is shown in the table 1.
The amino acid sequences of LBD-ALF and LBD-Q are as follows:
LBD-ALF：CTYNVKPDLQRFELYFLGTVTC(SEQ ID NO.1)；
LBD-Q：CTYKVKPKLQRFKLYFLGTVTC(SEQ ID NO.2)。
TABLE 1 analysis of physicochemical Properties of LBD-Q short peptides
Figure BDA0003097042790000091
EXAMPLE 2 Artificial Synthesis of short peptide LBD-Q short peptide
The LBD short peptide which is designed by engineering is synthesized. The synthesis method is based on solid phase chemical synthesis, the synthesis direction is from the C end to the N end of the LBD-Q short peptide, acetylation modification is carried out on the N end, amidation modification is carried out on the C end, and the end closure is formed to play a role in enhancing the stability of the synthesized polypeptide. A disulfide bond is formed between two cysteines, the synthesized LBD-Q short peptide is purified by reversed phase high performance liquid chromatography and the purity of the synthesized LBD-Q short peptide is detected, mass spectrometry is used for identifying the LBD-Q short peptide, the results are respectively shown in figure 1 and figure 2, the purity of the LBD-Q short peptide provided by the embodiment is 95.97% according to the calculation of figure 1, and the molecular mass of the LBD-Q short peptide provided by the embodiment is 2886.52 as can be seen from the mass spectrometry detection result shown in figure 2.
Example 3 Capacity of short peptide LBD-Q to clear WSSV Virus
In order to confirm the antiviral activity of the LBD-Q short peptide, WSSV virus solution is firstly prepared, the absolute quantitative detection is carried out on the virus copy number by using a Taqman probe method, the mass spectrum detection result of the synthesized TaqMan probe-WSSV32706 is shown in figure 3, and the DNA sequence is correct and the purity meets the requirement.
Then the LBD-Q short peptide and WSSV virus solution are incubated together, and the steps are as follows:
(1) the concentration of WSSV virus solution was adjusted to 2.5X 10 with sterilized physiological saline4copy/μL；
(2) Dissolving the short peptide LBD-Q to a concentration of 80 mu M;
(3) and incubating the WSSV virus solution and the short peptide LBD-Q solution with the same volume for 1h at room temperature by a shaking table to obtain a mixed solution to be injected, and performing subsequent injection experiments in time, wherein the mixed solution is named as a WSSV + LBD-Q group.
Comparative example 1
This comparative example differs from example 3 only in that the short peptide LBD-Q solution was replaced with physiological saline and designated as WSSV + sterilasine group.
Comparative example 2
This comparative example differs from example 3 only in that the short peptide LBD-Q solution was replaced with a pGFP solution, which was designated as WSSV + pGFP group.
Comparative example 3
This comparative example differs from comparative example 1 only in that no virus was added, and was designated as the steriesaline group.
Experimental example 1 injection experiment
Dividing 400 Penaeus japonicus into 10 groups of 40 per group at random, grouping according to example 3 and comparative examples 1-3, injecting 50 μ L of prepared mixture to be injected into the second abdominal node of each group of Penaeus japonicus, and finally respectively containing 1.25 × 10 of WSSV positive control, negative control and each tail of the group of Penaeus japonicus6copy WSSV. The results of plotting the effect of mortality of the penaeus japonicus obtained in example 3 and comparative examples 1 to 3 are shown in fig. 4, and it can be seen that the cumulative mortality of the experimental group after incubation of WSSV and LBD-Q is much lower than the cumulative mortality of the blank control group and the negative control group.
The curve drawing method is that after the WSSV group is injected, the death number is counted every 4 hours, and dead shrimps are fished out in time. Statistical significance tests were performed on differences in mortality between the experimental and control groups using the Kaplan-Meier plot (log-rank χ 2test) method in GraphPad Prism 8 software, and the cumulative mortality for each group was counted and plotted to obtain fig. 4.
Experimental example 2
After the injection in the WSSV group, 4 muscle samples of each of the Penaeus japonicus were sampled in ethanol at 96 hours, and DNA extraction of the samples was performed by a marine animal tissue genome DNA extraction kit. The absolute quantification procedure for the detection of viral copies of WSSV is described in Qiu et al (Qiu W, Zhang S, Chen Y, et al, Litopenaeus vannamei NF-. kappa.B is required for WSSV reproducibility [ J ]. development & Comparative Immunology,2014,45(1):156-162) with appropriate modifications:
(1) primers WSSV32678-F, shown as SEQ ID No.3, and WSSV32753-R, shown as SEQ ID No.4 (see Table 2) were designed based on the sequence of WSSV IE1(WSSV069) for PCR amplification, and the desired fragment was constructed in pMD19-T plasmid.
(2) And (4) carrying out subsequent recombinant plasmid extraction on the bacterial liquid with the correct sequencing.
(3) The sequence of the probe WSSV32706 is designed as shown in SEQ ID No.5 (see Table 2), the purification mode of the probe adopts HPLC, and 6-FAM at the 5 'end and TAMRA at the 3' end are modified.
TABLE 2 Probe and primer sequences
Figure BDA0003097042790000111
(4) Diluting the recombinant plasmid according to a gradient of 10 times to be used as a standard solution template, adding the extracted DNA of the prawn muscle sample into a system according to the mass of 100ng to be used as a sample template, and using WSSV32678-F and WSSV32753-R as primers to detect the copy number of WSSV in the muscles of the Japanese marsupenaeus japonicus after challenge by an absolute quantitative method, wherein the specific reaction system and the operation program are as follows:
TABLE 3 PCR reaction System
Figure BDA0003097042790000112
Figure BDA0003097042790000121
The reaction procedure is as follows: pre-denaturation at 95 ℃ for 30s, followed by extension at 95 ℃ for 5s, followed by fluorescence acquisition at 60 ℃ for 34s, and the extension and fluorescence acquisition steps were repeated for 40 cycles.
Three replicates per sample were performed. Obtaining the slope and R according to the standard curve of the sample displayed by the software2Values and amplification efficiency, and according to the formula: copy number mass/DNA molecular weight × 6.0 × 1023The copy number is obtained. The number of virus copies per 0.1. mu.g of shrimp tissue DNA was obtained.
Meanwhile, the copy numbers of the WSSV viruses obtained in the example 3 and the comparative examples 1 and 2 are detected after 96h of infection, and the results are shown in FIG. 5, and after 96h of infection, by absolutely and quantitatively detecting the copy numbers of the viruses, the WSSV copy number of the shrimps in the treatment group of the example 3 is remarkably reduced (p is less than 0.05) and the WSSV resistance is enhanced compared with the negative control group of the comparative example 2 and the blank control group of the comparative example 1.
Example 4
The embodiment provides a prawn feed which comprises a main material, an auxiliary material and an additive, and is composed of the following raw materials in parts by mass:
the main material comprises 20 parts of soybean meal and 10 parts of peanut meal;
the auxiliary materials comprise 25 parts of starch slurry, 0.05 part of ammonium sulfate and 0.05 part of sodium sulfite;
the additive comprises 2.5 parts of compound mineral substance, 1.5 parts of fish oil, 0.15 part of compound vitamin and 3 parts of short peptide LBD-Q coupled with outer membrane protein;
the molar ratio of the outer membrane protein to the short peptide LBD-Q is 1: 1.
Example 5
The embodiment provides a medicated bath liquid which comprises the following components in parts by weight:
10 parts of short peptide LBD-Q coupled with outer membrane protein, 5 parts of arginine, 4001 parts of polyethylene glycol, 801 parts of tween, 0.5 part of EDTA-2Na, 0.5 part of sodium sulfite, 5 parts of glycerol and 5 parts of ethanol, supplementing 100 parts of water for injection and adjusting the pH value to be about 7.0.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
SEQUENCE LISTING
<110> Zhuhai school district of Beijing university of teachers and professions
Application of <120> short peptide LBD-Q in preparation of antiviral infection product, feed and medicament
<160> 5
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> PRT
<213> marsupenaeus japonicum (marsupenaeus japonicus)
<400> 1
Cys Thr Tyr Asn Val Lys Pro Asp Leu Gln Arg Phe Glu Leu Tyr Phe
1 5 10 15
Leu Gly Thr Val Thr Cys
20
<210> 2
<211> 22
<212> PRT
<213> Artificial sequence
<400> 2
Cys Thr Tyr Lys Val Lys Pro Lys Leu Gln Arg Phe Lys Leu Tyr Phe
1 5 10 15
Leu Gly Thr Val Thr Cys
20
<210> 3
<211> 27
<212> DNA
<213> Artificial sequence
<400> 3
tgttttctgt atgtaatgcg tgtaggt 27
<210> 4
<211> 18
<212> DNA
<213> Artificial sequence
<400> 4
cccactccat ggccttca 18
<210> 5
<211> 28
<212> DNA
<213> Artificial sequence
<400> 5
caagtaccca ggcccagtgt catacgtt 28

Claims (20)

1. The application of the short peptide LBD-Q in the preparation of products for resisting WSSV virus infection is characterized in that the amino acid sequence of the short peptide LBD-Q is shown as SEQ ID No. 2.
2. Use according to claim 1, wherein the two cysteine residues in the sequence shown in SEQ ID No.2 are linked by a disulfide bond.
3. The use according to claim 1, wherein the sequence of SEQ ID No.2 has protective modifications at both ends.
4. The use of claim 3, wherein the protective modification comprises a chemical modification.
5. The use according to claim 3 or 4, wherein the amino terminus of the sequence of SEQ ID No.2 has an acylating modification.
6. The use according to claim 3 or 4, wherein the carboxy terminus of the sequence of SEQ ID No.2 has an amidation modification.
7. The use according to claim 1, wherein the product against viral infections comprises a medicament or a feed.
8. A feed, which is characterized by comprising a main material, an auxiliary material and an additive, wherein the additive comprises the short peptide LBD-Q in the application of any one of claims 1 to 7.
9. The feed of claim 8, wherein the main material comprises at least one of seeds, bran or meal.
10. The feed of claim 8, wherein the adjuvant comprises at least one of a binder, a mold inhibitor, or an antioxidant.
11. The feed of claim 8, wherein the additive further comprises at least one of a mineral element based additive, an animal derived protein based additive, a vitamin based additive, an amino acid based additive, an allicin based additive, or a growth factor additive.
12. The feed of any one of claims 8 to 11, wherein the short peptide LBD-Q is coupled to an outer membrane protein.
13. The feed of claim 12, wherein the outer membrane protein is derived from salmonella.
14. The feed of claim 13, wherein the feed comprises a shrimp farming feed.
15. The feed of claim 14, wherein the shrimp comprises marsupenaeus japonicus.
16. A medicament, characterized by comprising the short peptide LBD-Q and pharmaceutically acceptable auxiliary materials for the use according to any one of claims 1 to 7.
17. The pharmaceutical of claim 16, wherein the short peptide LBD-Q is coupled to an outer membrane protein.
18. The medicament of claim 17, wherein said outer membrane protein is derived from salmonella.
19. The medicament of any one of claims 16 to 18, wherein the medicament is in a form selected from the group consisting of a suspension, a granule, a spray, an emulsion, and a medicated bath.
20. The medicament of claim 19, wherein the auxiliary materials of the medicament comprise a cosolvent, an antioxidant, a pH modifier, a solvent, and a penetrant.
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