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Scatophagus argus luteinizing hormone (LH) gene, scatophagus argus LH recombinant protein and application

A technology of luteinizing hormone and recombinant protein, applied in the field of genetic engineering, can solve problems such as the lack of in vitro culture models, restrictions on the study of sperm molecular mechanisms, and environmental damage to humans

Inactive Publication Date: 2017-02-22
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Problems existing in the existing technology: With the continuous expansion of the scale of aquaculture, in order to ensure the stable production mode of the quantity and ensure the efficiency of reproduction and growth, many drugs are put into the production industry, which will undoubtedly cause certain harm to the environment and the human body
Since the development of germ cells requires a strict special environment, the lack of in vitro culture models has restricted the study of the molecular mechanism of spermatogenesis for a long time.

Method used

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  • Scatophagus argus luteinizing hormone (LH) gene, scatophagus argus LH recombinant protein and application
  • Scatophagus argus luteinizing hormone (LH) gene, scatophagus argus LH recombinant protein and application
  • Scatophagus argus luteinizing hormone (LH) gene, scatophagus argus LH recombinant protein and application

Examples

Experimental program
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Effect test

Embodiment

[0088] 1. Induced expression of recombinant protein: Take 100ul of the frozen pET-LH / BL and pET-28a / BL stored at -80°C and inoculate 1ml of kanamycin-containing (Kana + ) in fresh LB liquid medium at 37°C for 1-3h with shaking. Then draw an appropriate amount of bacterial liquid and apply it on the Kanamycin-containing + ) on LB solid plate medium, cultured at 37°C for 10-14h. Pick a single clone and inoculate it in 1ml containing kanamycin (Kanamycin) + ) in a 1.5ml EP tube of LB liquid medium, at 37°C, shake culture for 8-10h to allow Escherichia coli to amplify in large quantities (grow to the logarithmic growth phase of Escherichia coli).

[0089] 2. After the PCR verification of the bacteria solution, inoculate the bacteria solution containing the target gene insert into 400ml of fresh (Kana + The final concentration is 50ug / m1) in LB liquid medium, and expanded at 37°C.

[0090] 3. Acquisition of recombinant crude protein: centrifuge the collected bacterial liquid at...

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Abstract

The invention discloses scatophagus argus luteinizing hormone (LH) gene, scatophagus argus LH recombinant protein and application, in particular to the scatophagus argus LH recombinant protein obtained from the scatophagus argus luteinizing hormone (LH) gene through inducible expression of a prokaryotic expression system, and application of the scatophagus argus LH recombinant protein; the cDNA sequence overall length of the scatophagus argus luteinizing hormone (LH) gene is obtained, and the scatophagus argus LH recombinant protein is also obtained through prokaryotic expression; the pure LH recombinant protein is obtained through purification of an affinity column, and can be applied to in-vitro injection of fish to promote gonad development of the fish, and further the gonad development of the fish can show synchronism.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a cDNA sequence of a money fish luteinizing hormone LH gene, and the purification and application of a money fish LH recombinant protein obtained by constructing a recombinant plasmid and inducing expression of a prokaryotic expression system. Background technique [0002] Scatophagus argus, also known as golden drum fish, belongs to Perciformes (Perciformes), Scatophagidae (Scatophagidae), and is distributed in the East my country Sea and South China Sea. It is a high-quality economic fish with ornamental value. Moneyfish is a euryalline fish, which is distributed in sea areas with extremely high salinity, and can grow and survive well in pure fresh water. It can be directly transferred from 35‰ sea water to fresh water without desalination and domestication. Its rare adaptability to salinity makes it very suitable as a material for studying the reproducti...

Claims

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Application Information

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IPC IPC(8): C12N15/16C12N15/70C07K14/59A61K9/08A61K38/24A61P15/00
CPCA61K9/0019A61K9/08A61K38/00C07K14/59C12N15/70
Inventor 王武张俊彬
Owner SHANGHAI OCEAN UNIV
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