Pseudosciaena crocea interleukin 2 gene promoter sequence and application thereof

A large yellow croaker and promoter technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problem of activity upregulation

Active Publication Date: 2022-04-08
THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immune stimulants LPS and Poly(I:C) have little effect on its activity, but T cell activators PHA, Con-A, CI, PMA, etc. can stimulate IL-2 gene promoter activity to be significantly up-regulated

Method used

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  • Pseudosciaena crocea interleukin 2 gene promoter sequence and application thereof
  • Pseudosciaena crocea interleukin 2 gene promoter sequence and application thereof
  • Pseudosciaena crocea interleukin 2 gene promoter sequence and application thereof

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Experimental program
Comparison scheme
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Embodiment Construction

[0021] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.

[0022] 1. Cloning of the IL-2 gene promoter of large yellow croaker

[0023] 1.1 The GenomeWalker Universal Kit kit from Clontech Company was used to construct the large yellow croaker genome single-digestion library with 4 restriction endonucleases, and the 4 DNA endonucleases used were DraI, EcoRV, PvuII and StuI.

[0024] The specific operation is:

[0025] 1.1.1 Genome purity analysis.

[0026] ① Take 1 μL of large yellow croaker genomic DNA and electrophoresis on 0.6% agarose gel.

[0027] ②Use the endonuclease Dra I to test the digestion, the reaction system is:

[0028]

[0029] 1.1.2 Enzyme digestion of genome.

[0030] ① Label five 1.5mL centrifuge tubes D1, D2, D3, D4 and a positive control.

[0031] ②The components of each endonuclease digestion system include:

[0032]

[0033] ③Vortex at low speed for 5-10s and centrifuge brie...

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Abstract

The invention relates to a larimichthys crocea interleukin 2 gene promoter sequence and application thereof, and provides a larimichthys crocea IL-2 gene promoter sequence, an activity analysis method of a larimichthys crocea IL-2 gene promoter and application of the larimichthys crocea IL-2 gene promoter. The method comprises the following steps: cloning a large yellow croaker IL-2 gene promoter by adopting a genome walking method, and providing a large yellow croaker IL-2 gene promoter sequence; reporter gene analysis experiments prove that the sequence has promoter activity, and the activity of the sequence is not influenced by immune stimulants LPS and Poly, but can be obviously up-regulated under the induction of T cell activators PHA, Con-A, CI, PMA and the like. Cloning and activity identification of the promoter provide a good experimental system for researching expression regulation and control of the IL-2 gene of the large yellow croaker, create conditions for constructing an expression vector by using the promoter to efficiently express an exogenous gene or applying the expression vector to transgenic fish construction, and have important theoretical and practical significance.

Description

technical field [0001] The present invention relates to an interleukin 2 gene promoter, in particular to a large yellow croaker interleukin 2 gene promoter sequence and application thereof. Background technique [0002] Interleukin 2 (Interleukin, IL-2) is a pleiotropic cytokine that regulates CD4 + The development and differentiation of T lymphocytes, the promotion of B lymphocyte proliferation and antibody production are considered to play an important role in regulating the immune response against pathogenic infection (Gaffen S, Liu KD, Overview of interleukin-2 function, production and clinical applications[J] , 2004, Cytokine, 28(3):109-123). In the previous study, the full-length cDNA sequence of interleukin 2 gene (IL-2) of large yellow croaker was cloned from the spleen tissue of large yellow croaker, and the recombinant expression and purification of its signal peptide fragment were completed in Pichia pastoris, confirming that it can The expression of cytokines a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85C12N15/53C12N15/12C12Q1/26G01N21/64
CPCY02A40/81
Inventor 敖敬群慕鹏飞汪玉华陈新华
Owner THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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