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Application of C-type lectin

A lectin, antibacterial technology, applied in the field of molecular biology, can solve the problem of no report of antiviral C-type lectin, and achieve the effect of improving the ability of antiviral and antibacterial infection

Inactive Publication Date: 2015-05-20
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently known vertebrate C-type lectins mainly act on bacteria and yeast, and no antiviral C-type lectins have been reported

Method used

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  • Application of C-type lectin
  • Application of C-type lectin
  • Application of C-type lectin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Preparation of C-type lectin recombinant protein

[0016] 1) Construction of plasmid pCTL1 expressing C-type lectin recombinant protein:

[0017] The C-type lectin gene sequence in the present invention has been reported (GenBank accession number XP_008329461.1). The cDNA of tongue sole was used as a template and PCR amplification was carried out with primers F1 and R1. The PCR conditions were: 94°C for 60s to pre-denature template DNA, then 94°C for 40s, 63°C for 60s, 65°C for 60s, 30 cycles. PCR products were purified with corresponding kits from Tiangen. The expression vector pET259 (see Hu YH, Zheng WW, Sun L. Identification and molecular analysis of a ferritin subunit from red drum (Sciaenops ocellatus) for the construction process of pET259. Fish Shellfish Immunol 2010; 28:678-86) with restriction endonuclease After digestion with enzyme SwaI, connect with the above purified PCR product with T4 DNA ligase, transform the connection solution into Escherichia coli...

Embodiment 2

[0025] Immunological application of recombinant C-type lectin

[0026] Step 1) Injection of lectin

[0027] Dilute the C-type lectin obtained in Example 1 above to 20 ug / ml in PBS, which is the lectin dilution. Twenty tongue soles (about 10.1 g in weight) were randomly divided into 4 groups, 5 in each group. These 4 groups are named A, B, C and D respectively. Each fish in groups A and C was injected with 50ul of the lectin dilution, and each fish in groups B and D (control group) was injected with 50ul of PBS.

[0028] The composition of the PBS is by weight percentage: 0.8% NaCl, 0.02% KCl, 0.358% Na 2 HPO 4 .12H 2 O, 0.024% NaH 2 PO 4 , and the balance is water.

[0029] Step 2) Bacteria and virus suspension preparation

[0030] Cultivation of Vibrio anguillarum C312 to OD in LB medium 600 0.8, then centrifuged (5000g, 4°C, 10min), collected the bacteria, suspended in PBS to a final concentration of 10 6 cfu / ml is Vibrio eelium suspension. The cytomegalovirus RB...

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Abstract

The invention belongs to the field of molecular biology and particularly relates to application of C-type lectin. The C-type lectin is used for preparing inhibitors for inhibiting bacterial and viral infection. The recombinant C-type lectin is used for preparing preparations for inhibiting bacterial and viral infection of fishes and is capable of enhancing the bacterium and virus resistance of fishes significantly.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to the application of fish C-type lectin. Background technique [0002] Lectins are a class of sugar-binding proteins widely found in animals and plants. Animal lectins are divided into several different subclasses, one of which is C-type lectin. All C-type lectins have a conserved carbohydrate-recognition domain (carbohydrate-recognition domain), which is involved in substrate binding. Most C-type lectins rely on calcium ions for substrate recognition. C-type lectins play a role in a variety of biological processes, including cell-cell interactions and cell signaling. Many C-type lectins can combine with sugars on the surface of pathogens to cause pathogen agglutination, thereby inducing a series of immune responses and promoting the removal of pathogens. Therefore, C-type lectin is an important natural immune defense factor. The currently known vertebrate C-type lectins mainl...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P31/04A61P31/12
Inventor 孙黎周泽军
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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