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Gene of growth inhibitor in three kinds rockfish and its application

A technology of somatostatin and grouper, which is applied in the field of genetic engineering, can solve the problems of slow growth of grouper fry, and achieve the effects of improving growth rate and important economic benefits.

Inactive Publication Date: 2005-03-23
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide three kinds of grouper somatostatin genes aiming at the problem of slow growth of grouper fry breeding at present

Method used

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  • Gene of growth inhibitor in three kinds rockfish and its application
  • Gene of growth inhibitor in three kinds rockfish and its application
  • Gene of growth inhibitor in three kinds rockfish and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: Synthesis of the somatostatin gene PSSI of the grouper

[0026] Firstly, the cloning of the PSS I middle fragment was carried out. Compare the PSS I sequences of goldfish and other fish and other species, design and synthesize the upstream degenerate primer S 1 f 1 : 5'TCT CCA CGCGKA TCC AGT GC3' and downstream specific primer S 1 R 1 : 5' GTG AAA GTT TTCCAG AAG AA3'. Use GeneRacer TM The first-strand cDNA of the hypothalamus of the oblique-banded grouper obtained from Kit (Invitrogen Company, the same below) was used as a template to amplify the middle fragment of PSS I. The PCR reaction conditions were: 94°C pre-denaturation for 3 minutes, and then cycled: 94°C for 50s, 56°C for 1min, 72°C for 90s, a total of 30 cycles, and the last cycle was extended at 72°C for 10 minutes. The amplified products were electrophoresed on 1.5% agarose gel, stained with ethidium bromide, and specific fragments were purified by gel recovery. The specific fragment was...

Embodiment 2

[0029] Embodiment 2: Synthesis of the somatostatin gene PSSII of the grouper

[0030] Synthetic primers were designed according to the PSS II sequence of fish and other species to amplify the 3' end sequence of PSS II. upstream primer S 2 f 2 : 5'C TTC TAY TGG AAG GGC TTC3', GeneRacer TM The 3'nested primer provided by Kit: 5'CGC TAC GTA ACGGCA TGA CAG TG3', the template and reaction conditions are the same as above. The methods of recovering specific fragments, connecting specific fragments with T vectors, transforming DH5α competent cells, screening recombinant plasmids, and sequencing DNA of recombinant plasmids are all the same as above. According to the sequencing results of the PSS II 3' end sequence, two primers were designed to clone the PSS II 5' end sequence, and S 2 UP 1 5'CAG AGG GCT GCTGAT TGG TC3' with GeneRacer TM The 5'first primer 5'CGACUG GAG CAC GAG GAC ACU GA3' amplification provided by Kit, the template is the same as above. Recover specific fragm...

Embodiment 3

[0031] Embodiment 3: Synthesis of the somatostatin gene PSSIII of the orange-banded grouper

[0032] Referring to the sequences of PSSIII such as goldfish and frogs, design and synthesize upstream primers to amplify the 3' end sequence of PSSIII. S 3 f 1 : 5'AgC SCC MTG CAA AAA CTT CT3', with GeneRacer TM The 3'nested primer provided by Kit: 5'CGC TAC GTA ACG GCATGA CAG TG3' is used for amplification, and the template is the same as above. The reaction conditions were pre-denaturation at 94°C for 3 minutes, and then cycled: 94°C for 50s, 50°C for 1min, and 72°C for 1min. A total of 30 cycles, and the last cycle was extended at 72°C for 10 min. The methods of recovering specific fragments, connecting specific fragments with T vectors, transforming DH5α competent cells, screening recombinant plasmids, and sequencing DNA of recombinant plasmids are all the same as above. Synthetic primers were designed according to the results of PSSIII 3' end sequence determination to ampl...

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Abstract

The invention discloses three kinds of rockfish somatostatin genes, which are a gene fragment with rockfish somatostatin gene cDNA sequence, obtained by using rockfish hypothalamic total RNA as a template and by the RT-PCR and Gene RACER methods; the three genes has an important application to the rockfish growth.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to three genes. Background technique [0002] Somatostatin (somatostatin, SRIF or SS) was first isolated from sheep hypothalamus by Branzeau et al. in 1973. Existing studies have found that somatostatin or structurally related antigens exist in the nervous system and digestive tract of various vertebrates and invertebrates, and they regulate various physiological processes in the body, including growth, development and metabolism, and have been identified as a Multifunctional peptide widely distributed in the nervous system and peripheral tissues. [0003] The research on the physiological function of fish SRIF mainly focuses on the regulation of hormones related to pancreas development and metabolism, as well as the regulation of pituitary GH secretion. Rainbow trout gene I and gene II products (SRIF-14 and SRIF-25) can reduce blood insulin and glucagon lev...

Claims

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Application Information

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IPC IPC(8): A23K20/00A23K20/142A23K20/147C07K14/46C12N15/12
Inventor 李文笙叶星林浩然
Owner SUN YAT SEN UNIV
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