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Freshwater shrimp vitellogenin Vg gene, encoding protein and application of freshwater shrimp vitellogenin Vg gene

A vitellogenin, green shrimp technology, applied in the fields of biotechnology and developmental regulation, can solve the problems of Vg gene silencing and expression that have not yet been reported, and achieve the effect of delaying time

Inactive Publication Date: 2014-12-10
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the silent expression of Vg gene in crustaceans

Method used

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  • Freshwater shrimp vitellogenin Vg gene, encoding protein and application of freshwater shrimp vitellogenin Vg gene
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  • Freshwater shrimp vitellogenin Vg gene, encoding protein and application of freshwater shrimp vitellogenin Vg gene

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Embodiment 1

[0033] Embodiment 1: Obtaining the full-length cDNA of Vg gene of freshwater shrimp

[0034] Extraction of total RNA: Select the livers of 3 to 4 mature female shrimps, quickly put them into a pre-cooled mortar filled with liquid nitrogen, and grind them rapidly. Total RNA was extracted using RNAiso Plus extraction reagent from Takara Company combined with the traditional phenolic extraction method, and the extraction method was referred to the instruction manual. The quality of the RNA was detected by agarose gel electrophoresis, and the OD260 / 280 ratio of the sample was analyzed by a spectrophotometer to be between 1.8 and 2.0, and the concentration of the RNA was determined.

[0035] cDNA first-strand synthesis: refer to the Takara M-MLV reverse transcription kit to synthesize the first-strand cDNA.

[0036] Cloning of the full-length cDNA sequence of the freshwater shrimp Vg gene:

[0037] According to the obtained full-length cDNA sequence of Vg gene (Macrobrachium ro...

Embodiment 2

[0055] Embodiment 2: the synthesis of green shrimp Vg gene dsRNA

[0056] Based on the nucleotide sequence of SEQ ID NO.1, use NCBI online dsRNA primer design software (http: / / www.flyrnai.org / cgi-bin / RNAi_find_primers.pl) to design and prepare double-stranded RNA within the open reading frame Primers (SEQ ID NO.3-SEQ ID NO.4), according to Transcript AidTM T7 High Yield Transcription kit (Fermentas, Inc., USA) kit instructions for in vitro transcription to synthesize dsRNA. The prepared dsRNA was dissolved in DEPC water, its concentration was measured and its purity was detected on 1.5% agarose gel. The above dsRNA solution was injected into the pericardial cavity of freshwater shrimp at a dose of 4 μg / g. The control group was injected with the same volume of DEPC water, and the female shrimp used in the experiment were all in the early stage of ovarian development (just after ovulation). Samples (N ≥ 3) were collected on days 1, 3, 5, 7, 9, 11, 13, 15, and 17 post-injecti...

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Abstract

The invention discloses a freshwater shrimp vitellogenin Vg gene, an encoding protein and an application of the freshwater shrimp vitellogenin Vg gene. The full-length cDNA sequence of the Vg gene is cloned from hepatopancreas of the freshwater shrimp by a primer group provided by the invention, and the expression of a vitellogenin gene in the ovary and the liver can be silenced after the dsRNA of the gene is injected to the pericardial cavity of a female freshwater shrimp, so that the ovary development of the freshwater shrimp is remarkably delayed. The invention provides a new thought and an effective means for solving the sexual precocity phenomenon of the freshwater shrimp and culturing good varieties.

Description

[0001] technical field [0002] The invention discloses a green shrimp vitellogenin Vg gene, encoded protein and its application, and relates to the field of biotechnology and developmental regulation, in particular to the cloning of the green shrimp vitellogenin gene, and to using the gene dsRNA to regulate the development of the green shrimp ovary speed. [0003] Background technique [0004] Freshwater shrimp, scientific name Japanese river prawn ( Macrobrachium nipponense ), commonly known as river shrimp, belongs to the order Decapoda, the family Palaemonidae, and the genus Macrobrachium. It is widely distributed in waters all over my country. freshwater cultured shrimp. According to the "China Fishery Yearbook" in 2012, the annual output of cultured freshwater shrimp in the country exceeds 230,000 tons, and the annual output value exceeds 15 billion yuan. It is one of the most promising species in the freshwater aquaculture industry. In recent years, with the shar...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12N15/113A01K67/033
CPCA01K67/0338A01K2207/05C07K14/43509C12N15/113C12N15/89
Inventor 白鸿坤傅洪拓吴滟乔慧李法君张文宜龚永生颜跃弟金舒博
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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