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Epinephelus coioides SRY box relative gene SOX and its use

A technology for oblique banded grouper and grouper, which is applied in the field of genetic engineering and can solve the problem of not finding the SRY box gene and the like

Inactive Publication Date: 2005-03-02
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fish, a single SRY box gene that determines sex differentiation has not been found

Method used

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  • Epinephelus coioides SRY box relative gene SOX and its use
  • Epinephelus coioides SRY box relative gene SOX and its use
  • Epinephelus coioides SRY box relative gene SOX and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Obtaining of Sox Gene

[0017] (1) Sampling: 4-year-old grouper was purchased from the marine aquaculture base of Daya Bay Aquaculture Center in Guangdong Province. The fish body was dissected, the gonads were taken out, some gonad samples were frozen in liquid nitrogen, and the other part of the gonad samples were fixed with Bouin’s fixative solution, paraffin sectioned, and tissue section observation revealed that the gonads were stage III early ovaries, such as figure 1 shown.

[0018] (2) RNA extraction: total gonad RNA was extracted with Trizol Reagent, electrophoresed on 1% agarose gel, and RNA quality was checked. Electrophoresis showed that 18S and 28S ribosomal RNA bands were clear, indicating that the extracted ovarian RNA was of high quality. Such as figure 2 As shown, the positions of 18S and 28S ribosomal RNAs have been marked in the figure.

[0019] (3) cDNA library construction: take 1 μg gonad total RNA and apply SMARTT TM The cDNA libra...

Embodiment 2

[0021] Application of embodiment 2 Sox gene

[0022] (1) Tissue distribution of Sox gene expression in the grouper: extract total RNA from different tissues of the grouper, use RNase-free DNaseI to remove the contamination of genomic DNA that may exist therein, then take 0.5ug RNA, One-step reverse transcription PCR (One-stepRT-PCR) analysis kit (Invitrogen) was used to analyze the expression of Sox gene in different tissues, and the primers used were QOSOXF1 (5'GAT CAT GGT TCA ACA CAC GGA AC3') and QOSOXR1(5 'ACC GCT TCC CGA GCC GTT TAG AG3'), the number of PCR cycles is 39, and the size of the amplified product is 220bp. The results of one-step RT-PCR showed that the expression level of Sox gene in the grouper was higher in the gonad than in other tissues, such as Image 6 Shown, M, 100bp DNA molecular weight standard; 1 is the negative control (template is water); 2 is the gonad of the 4-year-old grouper (early stage III ovary); 3-10 are the gonads of the 2-year-old groupe...

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Abstract

The present invention relates to Epinephelus coioides SRY box relative gene Sox and its application. The present invention finds the gene Sox from the sex gland of Epinephelus coioides and the gene has important application in sex gland development and sex reversal of Epinephelus coioides.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to an SRY box-related gene Sox of a grouper and its application. Background technique [0002] Slanted grouper (Epinephelus coioides) belongs to the order Perciformes, the family Apiidae, the subfamily of groupers, and the genus Grouper. Its meat is delicious and nutritious, and it is the first-class delicacy on the dining table. Because of its fast growth and high and stable market price, it is an important economic fish for marine aquaculture in coastal areas of my country. The oblique-banded grouper is a kind of hermaphroditic fish with prefemininity. It is female at the beginning, and sex reversal occurs when the body length is 250-400mm. The male needs to be about 6 years old to mature, and it is often difficult to obtain simultaneously during artificial reproduction. The problem of functional male and female broodstock makes seed production very difficult. Therefore, it i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12
Inventor 张为民林丹张利红张勇
Owner SUN YAT SEN UNIV
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