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Prawn protein with antiviral activity and application of prawn protein

A technology of antiviral activity and prawn protein, which can be applied in antiviral agents, medical preparations containing active ingredients, applications, etc., and can solve problems such as applications that have not been reported

Active Publication Date: 2015-01-07
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching, its sequence and its application in antiviral infection or treatment of shrimp, crab and other animals have not been reported

Method used

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  • Prawn protein with antiviral activity and application of prawn protein
  • Prawn protein with antiviral activity and application of prawn protein
  • Prawn protein with antiviral activity and application of prawn protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 Contains the acquisition of the characteristic sequence protein in prawns

[0035] Isolate the gill tissue of prawns, extract tissue total RNA according to the instructions of the TRIzon kit (Clontech), reverse transcribe into cDNA according to the instructions of the Super SMART PCR cDNA synthesis Kit user manual (Clontech), and provide SEQ ID NO: 2 according to the present invention Design expression primers:

[0036] ExF: 5'-TACTCAGGATCCATCAGCCGGCCAGC-3';

[0037] ExR: 5'-TACTCACTCGAGCTGGTTGGCTTTCTTGA-3', add BamH I and Xho I restriction sites. The coding sequence of the protein was obtained by PCR amplification.

[0038] Digest the coding sequence recovered from the gel with the prepared pET30a(+) vector, respectively. The digested PCR fragments are recovered by the product, and the digested vector plasmid is recovered by gel recovery to recover the target fragment and plasmid respectively. The recovered target fragment was ligated with the vector, ...

Embodiment 2

[0039] The mensuration of embodiment 2 active protein reduction hydrogen peroxide activity

[0040] Catalase detection kit (S0051) was purchased from Beyond Biotechnology Research Institute. According to the kit instructions, a standard curve is drawn first, and then the activity of the active protein of the present invention is determined. Negative controls used BSA (bovine serum albumin) and other proteins of the same protein expression system.

[0041] (1) Standard curve drawing

[0042] Take 0, 12.5, 25, 50 and 75μL of 5mM H 2 o 2 Put the solution into a 5mL plastic centrifuge tube, add hydrogen peroxide detection buffer to a final volume of 100μL, and mix well. Take 4 μL each and add to a well in a 96-well plate. Add 200 μL of chromogenic working solution. Determination of A after incubation at 25°C for 15 min 520 .

[0043] (2) Sample determination

[0044] Table 1: Enzyme activity detection system

[0045]

[0046] Referring to the table above, take 10 μL o...

Embodiment 3

[0051] The antiviral effect of embodiment 3 active protein in prawn body

[0052] In this example, the white spot syndrome virus (WSSV), an important pathogenic virus of prawns, was selected to study the antiviral effect. This virus is a double-stranded DNA virus, which can infect a variety of crustaceans. It is an important pathogenic virus for marine animals. At present, there is no specific drug for its prevention and treatment.

[0053] Buy healthy Japanese prawns (10-15g / only) from the seafood market and keep them in the aquarium. Three prawns were randomly selected to extract the genomes of the gill tissues. Conventional PCR method was used to confirm that there was no virus infection. After feeding for 2-3 days to adapt to the environment, the experiment began. The prawns were divided into groups of 20 in each group, and the virus was injected respectively (10 6 copy number), active protein of the present invention (50μg)+virus (10 6 copy number) and control protein ...

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Abstract

The invention discloses prawn protein with antiviral activity. The prawn protein has an amino acid sequence as shown in SEQ ID NO.1. The invention further discloses nucleotide for coding the protein and an application of the protein in prawn and crab antiviral infection or treatment. The prawn protein disclosed by the invention further has the ability of reducing hydrogen peroxide, and also has the functions of lowering duplication amount of viruses in a living body, lowering a death rate of virus-contracted animals, and the like. The protein is expected to be developed into a pharmaceutic preparation for treating and preventing different virus infections in a plurality of living bodies.

Description

technical field [0001] The invention relates to an active protein and its application, in particular to a prawn protein with antiviral activity and the application of the protein in biological antiviral infection or treatment, belonging to the field of biotechnology. Background technique [0002] Viruses can be divided into two categories: true viruses and subviruses, among which true viruses can be further divided into two categories: DNA viruses and RNA viruses according to the type of viral nucleic acid. Viral diseases have the characteristics of long incubation period, complex transmission routes, violent onset, and lack of specific therapeutic drugs. They are currently important types of diseases that endanger the health of organisms. [0003] For vertebrates, the use of vaccines for immunization is an important method to prevent a variety of viral diseases. In addition, some antiviral drugs such as iodine deoxyuridine (herpesnet), vidarabine , acyclovir and interferon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/08C12N15/53A61K38/44A61P31/20
Inventor 康翠洁陈晓伟王金星
Owner SHANDONG UNIV
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