Paralichthys olivaceus SOCS3 gene and eukaryotic expression vector, expression system and heterologous expression method thereof

A technology of eukaryotic expression vector and expression system, which is applied in the field of flounder SOCS3 gene and its eukaryotic expression vector, can solve the problems such as lack of related content of flounder, and achieve the effect of easy transfection

Pending Publication Date: 2021-11-02
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although SOCS3 is well characterized in several mammals, there is little research on SOCS3 in flounder

Method used

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  • Paralichthys olivaceus SOCS3 gene and eukaryotic expression vector, expression system and heterologous expression method thereof
  • Paralichthys olivaceus SOCS3 gene and eukaryotic expression vector, expression system and heterologous expression method thereof
  • Paralichthys olivaceus SOCS3 gene and eukaryotic expression vector, expression system and heterologous expression method thereof

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Embodiment Construction

[0027] Of the accompanying drawings and the following the invention is further illustrated embodiment.

[0028] MATERIALS AND METHODS

[0029] 1.1 Materials

[0030] 1.1.1 vectors, cells, strain

[0031] Eukaryotic expression vector pEGFP-C1 purchased from Wuhan Miao Ling Biological Technology Co., HEK293T cells were purchased from Bai biological families, DH5α competent cells were purchased from engine Bioscience.

[0032] 1.1.2 Reagents

[0033] 2000 DNA Maker, restriction endonuclease EcoRⅠ, FD.Buffer, double distilled water, DNA Loading Buffer, DNA Gel Extraction Kit (column centrifugation), a small amount of high-purity plasmid DNA kit.

[0034] 1.1.3 The main experimental apparatus

[0035] Adjustable micro pipette (10μl, 200μl, 1000μl), 4 ° C refrigerator, the PCR instrument, the horizontal electrophoresis tank, centrifuge tubes (0.2ml, 1.5ml, 2ml), high-speed centrifuge, microcentrifuge, constant temperature water bath, the automatic gel imaging analyzer, clean bench, temp...

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Abstract

The invention discloses a paralichthys olivaceus SOCS3 gene as well as an eukaryotic expression vector, an expression system and a heterologous expression method of the paralichthys olivaceus SOCS3 gene. The nucleotide sequence of the paralichthys olivaceus SOCS3 gene is as shown in SEQ ID NO. 1. An upstream primer for amplifying the paralichthys olivaceus SOCS3 gene is as shown in SEQ ID NO.2, and a downstream primer is as shown in SEQ ID NO.3. On the basis of successfully cloning the paralichthys olivaceus SOCS3 gene, a eukaryotic expression vector pEGFP-C1-PoSOCS3 of the paralichthys olivaceus SOCS3 gene is further constructed, the eukaryotic expression vector pEGFP-C1-PoSOCS3 is successfully transfected into an HEK293T cell, subcellular localization of the paralichthys olivaceus SOCS3 gene in the cell is observed by a fluorescence inverted microscope, and basic data is provided for subsequent paralichthys olivaceus SOCS3 function research.

Description

Technical field [0001] The present invention relates to the field of molecular biology, more particularly to a gene SOCS3 flounder its eukaryotic expression vector, heterologous expression systems and expression method. Background technique [0002] Flounder (Paralichthysolivaceus), also referred to Paralichthys olivaceus, commonly known as dental films, flatfish, belonging Osteichthyes, Pleuronectiformes, paralichthyidae, is near the sea bottom living cold temperature fish, having a latent sand habit, as euryhaline fish. At the same time economic value and nutritional value flounder are high, with succulent fresh, nutritious, less miscellaneous thorn, fast growth, etc., are widely farmed in some Asian countries, such as China, Japan, the Korean Peninsula. Our flounder are distributed in the coastal farming land, which belongs to the Yellow Sea and Bohai large number of breeding flounder. With the rapid development of flounder aquaculture industry, labor intensive farming become ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/85C07K14/46
CPCC07K14/461C12N15/85C12N2800/107
Inventor 王光花张敏
Owner QINGDAO AGRI UNIV
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