Gene sequence and use of embryogenesis regulatory factor in hypophysis of rockfish

A technology for embryonic development and regulatory factors, applied in genetic engineering, plant genetic improvement, medical preparations containing active ingredients, etc., can solve problems such as cloning and separation

Inactive Publication Date: 2003-03-12
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

No genes regulating the embryonic development of grouper have been cloned yet

Method used

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  • Gene sequence and use of embryogenesis regulatory factor in hypophysis of rockfish
  • Gene sequence and use of embryogenesis regulatory factor in hypophysis of rockfish
  • Gene sequence and use of embryogenesis regulatory factor in hypophysis of rockfish

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Experimental program
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Embodiment Construction

[0022] 1. Extraction of RNA and construction of SMART cDNA plasmid library

[0023] The pituitary total RNA was extracted with SV Total RNA Isolation System (Promega), and the steps were as follows: take about 500 grams of grouper, anesthetized and bled, removed the hypothalamus by craniotomy, added 175 μl of extraction buffer solution, and homogenized thoroughly, Add 350 μl RNA dilution buffer, mix well, block at 70°C for 3 minutes, and centrifuge at 14,000×g for 10 minutes. Transfer the supernatant, add 200 μl 95% ethanol, pipette and mix well, transfer to the spin column, centrifuge at 14000×g for 1 min, add 600 μl RNA washing solution, and centrifuge at 14000×g for 1 min. Add 50 μl DNase I to digest at 20-25°C for 15 minutes, then add 200 μl DNase stop solution, and centrifuge at 14000×g for 1 minute. Add 600 μl of SV RNA washing solution, centrifuge at 14000×g for 1 min, then add 250 μl of RNA washing solution, and centrifuge at high speed for 2 min. Total RNA was elute...

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Abstract

A gene sequence of an embryonic development regulatory factor in the scapus of Epinephelus has 514 bp for cDNA length, 252 bp for open reading frame, 83 coding amino acids, 53 bp for 5' non-coding region, and 209 bp for 3' non-coding region. It plays the important role in early development stage, and can be used in artificial breeding.

Description

technical field [0001] The present invention relates to the gene sequence of an embryonic development regulatory factor in the grouper pituitary, more specifically to a kind of embryonic developmental regulatory factor 1 (Gastrula Specific embryonic regulator 1, gerl) sequence, amino acid sequence, spatiotemporal expression pattern, the present invention also relates to a kind of use. Background technique [0002] Grouper (E.coioides) is marine coral reef fish, belongs to Serranidae (Serranidae), grouper (Epinephelus), is a kind of rare seawater fish, the sales market is stable, and the price is expensive. It is one of the marine fishes with the most technical difficulties in artificial propagation and nursery. So far, the supply of cultured seedlings still depends on the wild natural seedlings. In recent years, large-scale artificial breeding has been carried out in Japan, Southeast Asian countries, my country's Taiwan, and the coast of South China. Wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K20/147A23K50/80A61K31/7088A61P15/08C07H21/04C07K14/435C12N15/12C12N15/63
Inventor 周莉桂建芳
Owner INST OF AQUATIC LIFE ACAD SINICA
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