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Eriocheir sinensis GnRH analogue, and preparation method and application thereof

A Chinese mitten crab, hormone-releasing technology, applied in the field of molecular biology and biology

Inactive Publication Date: 2014-07-09
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The combination of RP-HPLC, time-resolved fluorescence analysis (TR-FIA) and immunohistochemistry in Penaeus japonicus also confirmed the existence of GnRH analogs (AMANO, OKUMURA, OKUBO, et al.Biochemical analysis and immunohistochemical examination of a GnRH-like immunoreactive peptide in the central nervous system of a decapod crustacean, the kuruma prawn (Marsupenaeus japonicus) [J]. Zoolog Sci, 2009, 26(12): 840-845.), but so far there is no Related reports on separation and purification of GnRH

Method used

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  • Eriocheir sinensis GnRH analogue, and preparation method and application thereof
  • Eriocheir sinensis GnRH analogue, and preparation method and application thereof
  • Eriocheir sinensis GnRH analogue, and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1 Fractionation, immunoassay and tandem mass spectrometry analysis of GnRH analogues in Chinese mitten crab

[0042] 1 Materials and methods

[0043] 1.1 Experimental materials

[0044] Peptide isolation experiment: Eriocheir sinensis was purchased from a local farm in Shanghai, and female sexually mature individuals with healthy limbs, strong vitality, and a body weight of 100-150g were selected to be temporarily raised in the Key Open Laboratory of Aquatic Germplasm Resources and Utilization of the Ministry of Agriculture, Shanghai Ocean University , Build a shelter in the breeding box (50cm×60cm), oxygenate for 24 hours, change the water every other day, and feed the feed twice a day.

[0045] 1.2 Experimental method

[0046] 1.2.1 Preparation of crab brain crude extract

[0047] Select female sexually mature healthy individuals, quickly dissect the brain tissue on ice, add 10ml of 1wt‰ trifluoroacetic acid aqueous solution to every 50 crab brain tissues, ...

Embodiment 2

[0075] Example 2 The function of mitten crab gonadotropin-releasing hormone analogs

[0076] 1 The first stage of experiments to verify the activity of GnRH analogues

[0077] 1.1 Materials and methods

[0078] 11.1 Experimental materials

[0079] Eriocheir sinensis was purchased from a local farm in Shanghai. Female sexually mature individuals with sound limbs, strong vitality, and uniform body weight were selected for breeding in the Aquaculture Center of the School of Aquaculture and Life Sciences, Shanghai Ocean University. The recirculating aquaculture system was used for breeding. Each breeding box (70cm×50cm×40cm) built a shelter, fed feed twice a day, oxygenated 24 hours, regularly replaced the circulating water in the breeding system to keep the water clean.

[0080] 1.1.2 Synthetic peptides

[0081] Entrusted Shanghai Jier Biochemical Co., Ltd. to artificially synthesize GnRH according to the amino acid sequence of the GnRH analogue of Chinese mitten crab. The pur...

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Abstract

The invention relates to the fields of molecular biology and biological technology, and discloses Eriocheir sinensis GnRH (gonadotropin-releasing hormone) analogues with respective amino acid sequence of QSYHFSHGWKP and QAYHFSHGWKP . Compared the the amino acid sequences of the two subtypes of Eriocheir sinensis GnRH analogues with those of the GnRH of other species, the results show that the former and the latter have similar amino acid conservative sites; the the sequence structures of the Eriocheir sinensis GnRH analogues are similar to that of invertebrate GnRH; and the Eriocheir sinensis GnRH analogues have similar functions as vertebrate GnRH, can stimulate breakdown of oocyte germinal vesicle and have distinct stimulative effect on crab oocyte maturation and important application value for artificial ripening of the Eriocheir sinensis.

Description

technical field [0001] The invention relates to the fields of molecular biology and biotechnology, in particular to a Chinese mitten crab gonadotropin-releasing hormone analogue and a preparation method and application thereof. Background technique [0002] Gonadotropin-releasing hormone (GnRH) of vertebrates is a neuropeptide hormone composed of ten amino acids secreted by the hypothalamus. GnRH, as a key information molecule of the hypothalamus-pituitary-gonad axis, binds to the GnRH specific receptor in the pituitary gland , thereby stimulating the production and release of gonadotropin (GSH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH), stimulating ovulation, increasing pregnancy rate and inhibiting ovarian development disorders, playing a role in regulating reproductive activities. In the 1970s, researchers began to study GnRH. In 1971, Schally et al. purified a hormone from pig hypothalamus that could promote the release of LH. After 6 years of res...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/23C07K1/16A01K61/00
CPCY02A40/81
Inventor 丘高峰王承志李真宋亚楠
Owner SHANGHAI OCEAN UNIV
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