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Epinephelus anti-transgenosis protein and nucleotide sequence and use thereof

A grouper and nucleotide technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of affecting the normal development of fish, labor and time-consuming, weakening, etc., and achieve the effect of enhancing fecundity and artificially controlling sex

Inactive Publication Date: 2005-07-20
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, it is not only labor-intensive and time-consuming, but also the conversion effect is poor. After sex conversion, the sperm vitality of male fish is not strong, and after the drug is stopped, sex reversion problems will occur.
Implantation of hormones will cause the fish to not eat or eat less, which will affect the normal development of the fish

Method used

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  • Epinephelus anti-transgenosis protein and nucleotide sequence and use thereof
  • Epinephelus anti-transgenosis protein and nucleotide sequence and use thereof
  • Epinephelus anti-transgenosis protein and nucleotide sequence and use thereof

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Experimental program
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Effect test

Embodiment Construction

[0054] 1. Extraction of RNA and construction of SMART cDNA plasmid library

[0055] The total RNA of the hypothalamus was extracted with SV Total RNA Isolation System (Promega), and the steps were as follows: take about 15 catties of grouper, anesthetized and bled, removed the hypothalamus by craniotomy, added 175 μl of extraction buffer solution, and homogenized fully , 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 14 000×g for 1 min, add 600 μl RNA washing solution, and centrifuge at 14 000×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 14 000×g for 1 minute. Add 600 μl SVRNA washing solution, centrifuge at 14 000×g for 1 min, then add 250 μl RNA washing solution, and centrifuge at high speed for 2 min. Total RNA was el...

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Abstract

The present invention discloses one kind of rockfish sex reversal relative gene sequence, vector and recombinant strain containing the gene and their use. The gene has cDNA of 608 bp length, one open reading frame of 252 bp (54-305), 83 coded amino acids, 5' non-coding region of 53 bp length and starting at one ANNATG motif included inside vertebrate initiator codon, and 3' non-coding region of 303 bp length and with polypeptide tailing signal (AAUAAA) and polypeptide tail. The vector and recombinant strain containing the gene and protoyon expressed fusion protein are used in propagation regulation, sex gland differentiation and sex conversion of rockfish and the feed additive preparation.

Description

technical field [0001] The present invention relates to a sequence of a grouper sex inversion related gene, more specifically to a sequence of a sex inversion related factor 1 (sex inversion related factor 1, sirf1) screened from a SMART cDNA library of a grouper pituitary , amino acid sequence, and the present invention also relates to the use of the carrier and recombinant strain of the gene. 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 S...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K20/147A23K50/80A61K38/17A61P15/00C07H21/00C07K14/435C12N15/12C12N15/62C12N15/63
Inventor 汪洋周莉桂建芳
Owner INST OF AQUATIC LIFE ACAD SINICA
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