Immune application of fish B cell activating factor

A technology of activating factors and B cells, applied in the field of molecular biology, can solve the problems of less research on fish BAFF and unknown functions, and achieve the effect of improving the immune protection effect

Inactive Publication Date: 2015-07-22
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

BAFF in fish is currently poorly studied and its function is almost completely unknown

Method used

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  • Immune application of fish B cell activating factor
  • Immune application of fish B cell activating factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Construction of the expression plasmid pBAFF of BAFF:

[0020] The sequence of BAFF of the present invention has been published (GenBank accession No. XM_008328682). BAFF consists of 261 amino acids and has a conserved TNF (tissue necrosis factor) domain. 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: pre-denatured template DNA at 94°C for 60s, then 94°C for 40s, 51°C for 60s, 72°C for 60s, after 5 cycles, changed to 94°C for 40s, 58°C for 60s, 72°C for 60s, 30 cycles. The PCR product was purified with the Tiangen DNA Product Purification Kit and ligated with the PCR cloning vector pBS-T (purchased from "Tiangen Biochemical Technology Co., Ltd.", Beijing) at room temperature for 2-4 hours, and the ligation mixture was transformed into Escherichia coli DH5α After containing 100 μg / ml anka penicillin (Ap), 40 μg / ml 5-bromo-4-chloro-3-indole-β-D-lactoside (X-gal) and 24 μg / ml isopropyl...

Embodiment 2

[0023] Immunization Application of BAFF Expression Plasmid pBAFF

[0024] Step 1) Immunization of tongue sole

[0025] The Edwardsiella tarda DNA vaccine plasmid pCEsa1 (see Sun Y, Liu C, Sun L. Construction and analysis of the immune effect of an Edwardsiella tarda DNA vaccine encoding a D15-like surface antigen. Fish Shellfish Immunol. 2011 for its construction process; 30:273–9.) and the pBAFF of the above-mentioned Example 1 were diluted to 400ug / ml in PBS, and the two were mixed in equal volumes to obtain the pCEsa1+pBAFF mixture. Dilute pCEsa1 and pBAFF to 200ug / ml in PBS respectively, namely pCEsa1 dilution and pBAFF dilution. 140 tongue sole (about 14.5g in weight) were randomly divided into 4 groups, 35 in each group. These 4 groups are named A, B, C and D respectively. Each fish in group A was injected intramuscularly with 0.1ml pCEsa1 dilution, each fish in group B was injected intramuscularly with 0.1ml pBAFF dilution, each fish in group C was injected intramusc...

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Abstract

The invention relates to the field of molecular biology, in particular to immune application of Cynoglossus semilaevis B cell activating factor (BAFF). The Cynoglossus semilaevis B cell activating factor is applied as a vaccine enhancement factor. The BAFF expression plasmid involved in the invention can significantly enhance the immunoprotection effect of DNA vaccines.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to the immunological application of B cell activating factor of tongue sole semismoothus. Background technique [0002] B cell activating factor (BAFF), belonging to the tumor necrosis factor (Tumor Necrosis Factor, TNF) family, is a typical type II transmembrane protein with a molecular weight of about 31kDa, composed of cytoplasmic region, transmembrane region and Composition of extracellular domains. In mammals, BAFF plays an important role in humoral immunity. It can bind and stimulate B lymphocytes and secrete immunoglobulins, and it also has a certain regulatory effect on the function of T cells. BAFF combined with its three receptors BAFF-R, TACI and BCMA can activate NF-κB, mTOR, p38, JNK and other signaling pathways, maintain the survival, differentiation and maturation of immune cells, and have anti-apoptotic effects. BAFF plays a key regulatory role in the development o...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K48/00C12N15/85C12N15/28A61P37/04
Inventor 孙黎孙云迟恒
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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