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Application of Vibrio alginolyticus-OmpA as animal immunity regulator

A technology of vibrio alginolyticus and animal immunity, which is applied in the direction of medical preparations containing active ingredients, drug combinations, antibody medical ingredients, etc.

Inactive Publication Date: 2008-06-25
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, before the present invention was published, there was no disclosure or report on the immunomodulatory function of Vibrio alginolyticus OmpA mentioned in this patent application

Method used

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  • Application of Vibrio alginolyticus-OmpA as animal immunity regulator
  • Application of Vibrio alginolyticus-OmpA as animal immunity regulator

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Preparation and Purification of Vibrio alginolyticus OmpA

[0014] In this example, the full-length coding sequence or fragment of Vibrio alginolyticus OmpA was constructed into the E. coli protein prokaryotic expression vector to express and purify the target protein.

[0015] The coding sequence of the Vibrio alginolyticus OmpA gene is as follows (shown in Sequence Listing 400):

[0016] 1 atgaaaaaac tagcagcggt aatttcagag actctactta tggcttcagc

[0017] 51 agcacaagca gaagtatacg ttggtggtaa gatgggtaag tcatggatgg

[0018] 101 aagacgcgtg tgtagcgggc caggcttgtg acaaagacga ttctactctt

[0019] 151 ggcgcttttg ttggttacga aatgaataaa tacttcgcag tagaagcggg

[0020] 201 tttcgacaac atcggtgatt ttaaccaaac ttctttcagt ggccacgtag

[0021] 251 aagcaatcac tcttgctcct aaatttagcc taccaatcac tgaagacatc

[0022] 301 gcactttacg gtaaagtggg tggcgcttac gtaatgtttg atggcaaaga

[0023] 351 tgattactct tacctaggcg cggctggtct tgaattcaac ctaagccaaa

[0024] 401 acgtaacagc tcgtgcggaa taccaaacac tga...

Embodiment 2

[0053] Preparation and Purification of Vibrio alginolyticus 1061 Protein

[0054] In this example, the full-length coding sequence or fragment of the Vibrio alginolyticus 1061 gene was constructed into the E. coli protein prokaryotic expression vector to express and purify the target protein.

[0055] The coding sequence of Vibrio alginolyticus 1061 gene is as follows (shown in Sequence Listing 400):

[0056] 1 atgcaactga acaaggttct aaaagggcta ctaatcgcgc ttcctgtaat

[0057] 51 ggctgttaca gcgtgtagtt caagcgacga cgctgcttct aacactggtg

[0058] 101 cagcaacaaa caacaatggc gcagtggaaa caacagtagc tgcgccaatt

[0059] 151 gatcaaagcg gtcagttgac agagcaagag ctaaaagagc aagcgctacg

[0060] 201 tgaaactcaa acaatctact ttgcttttga caactcaaca attgcagcag

[0061] 251 attacgaaga gatgctagcg gctcacgctt cttacctaag caagaaccca

[0062] 301 gcacttaacg taactatcga aggtcacgct gatgagcgcg gtacacctga

[0063] 351 gtacaacatt gctctaggtg agcgtcgtgc acaagcagta gcgaactacc

[0064] 401 tacaagcact aggtgttcaa g...

Embodiment 3

[0079] Active cross-immune protection of Vibrio alginolyticus OmpA to crucian carp

[0080] 1. Immunization of crucian carp: Feed the bought crucian carp for 1 week, and feed artificial compound feed twice a day in the morning and evening. There was no abnormal grouping after 1 week of domestication. Take the purified OmpA and mix it with an equal volume of complete adjuvant (lanolin: paraffin oil = 1:4 v / v, 30 mg / ml for BCG), and after repeated suction with a syringe, ultrasonic treatment for 2 minutes, that is, the prepared antigen, intraperitoneal Inject immunized crucian carp, 25 μg / tail, and the control group does not receive immunization. Injected twice, with an interval of 10 days each time, the experimental group was injected with incomplete adjuvant mixed with the purified protein of interest (12.5 μg / fish) for the second time.

[0081] 2. Evaluation of cross-immune protection: 10 days after the second immunization, the corresponding bacteria (Vibrio alginolyticus a...

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Abstract

The invention relates to a vibrio alginolyticus (E.coli) which reflects the immune regulating function of a recombinant outer membrane proteinA (outer membrane proteinA, OmpA). The invention can remarkably improve the resistance of pathogens of the vibrio alginolyticus, pseudomonas fluorescens, etc. after the crucian is immunized through recombining the outer membrane proteinA of the vibrio alginolyticus. The test of phagocytosis in vitro can obviously affect the mononuclear macrophage on phagocytic function of bacteria. After mixed immunity with vibrio alginolyticu, the invention obviously improves the tilter tests of 1061 protein antibodies, which provides escherichia coli OmpA which is provided with the functions of animal immune regulation and improves the effect of animal disease resistance ability.

Description

technical field [0001] The invention relates to the immune regulation function of an outer membrane protein A (outer membrane protein A, OmpA) expressed by Vibrio alginolyticus. Background technique [0002] Immunomodulators refer to a large class of drugs that can regulate, enhance, excite and restore the immune function of the body. In aquatic animal farming, it is a common measure to use various chemical drugs and antibiotics to control related diseases. However, long-term use of drugs not only obviously leads to the increase of drug resistance of pathogenic bacteria, but also the hazards of drug residues are increasingly apparent, which seriously affects the safety of aquatic products. . As a result, aquaculture animal immunomodulators came into being. [0003] Immunomodulators are also called immunopotentiators or immunostimulators. When used together with antigens, they can stimulate animal-specific immune mechanisms, and when used alone, they can stimulate non-speci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61P37/04
Inventor 李惠熊筱鹏王保成彭宣宪
Owner SUN YAT SEN UNIV