Freshwater shrimp opsin gene, protein encoded by same and application

A technology of gene encoding and opsin, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problem that opsin gene silencing expression has not been reported yet, and achieve the effect of reducing the expression of vitellogenin gene

Active Publication Date: 2018-05-18
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Description
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  • Application Information

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Problems solved by technology

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

Method used

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  • Freshwater shrimp opsin gene, protein encoded by same and application
  • Freshwater shrimp opsin gene, protein encoded by same and application
  • Freshwater shrimp opsin gene, protein encoded by same and application

Examples

Experimental program
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Embodiment 1

[0017] The acquisition of the full-length cDNA of embodiment 1 freshwater shrimp opsin gene

[0018] Total RNA extraction: select 3-4 mature female shrimp, aseptically dissect and remove the eye tissue, clip the eye stalk pigment in RNA preservation solution (TaKaRaBioInc.Japan), rinse and pre-treat, use TaKaRa Trizol kit Extract the total RNA of eye stalk of freshwater shrimp, and refer to the instruction manual of the kit for the specific operation steps.

[0019] The integrity of the total RNA was detected by 1.2% agarose gel electrophoresis, and the concentration and OD value of the extracted RNA were measured by a BioPhotometer ultraviolet spectrophotometer. Usually, the A260 / A280 was between 1.8 and 2.0, which met the experimental requirements.

[0020] First-strand cDNA synthesis: refer to the Takara M-MLV reverse transcription kit to synthesize the first-strand cDNA from the above total RNA.

[0021] Obtaining the full-length cDNA of the freshwater shrimp opsin gene: ...

Embodiment 2

[0031] The synthesis of embodiment 2 freshwater shrimp Opsin gene dsRNA

[0032] According to 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 strands within the open reading frame of the nucleotide sequence RNA primers (SEQ ID NO: 3-SEQ ID NO: 4), using the total cDNA of freshwater shrimp as a template, performed in vitro transcription to synthesize dsRNA according to the instructions of the TranscriptAidTMT7 High Yield Transcription kit (Fermentas, Inc., USA). The prepared dsRNA was dissolved in DEPC water, its purity was detected on 1.5% agarose gel and its concentration was measured by a spectrophotometer. Female freshwater shrimps in the early stages of ovarian development (just after ovulation) were equally divided into two groups (experimental group and control group). The experimental group was injected with dsRNA solution at a dose of 4ug / g, and the c...

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Abstract

The invention discloses a freshwater shrimp opsin gene, a protein encoded by the same and application. The nucleotide sequence of the gene is SEQ ID NO: 1, and the nucleotide sequence encoded by the gene is SEQ ID NO: 2. A molecular mechanism for regulation and control of key genes related to exogenous environmental factors to development of ovary of crustacean, and a theoretical basis is providedfor sexual precocity of freshwater shrimps; and (2), dsRNA which is synthesized according to the nucleotide sequence of the freshwater shrimp opsin gene is injected into the cardiocoelom of a freshwater shrimp, vitellogeningene expression in the ovary of the freshwater shrimp can be relieved effectively, and therefore, the freshwater shrimp opsin gene has quite important influences to developmentof the ovary of the freshwater shrimp.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to the cloning and application of a functional gene, specifically the cloning of the opsin gene of freshwater shrimp, and the cDNA of the gene is used as a template to synthesize dsRNA, and the dsRNA is used to affect the yolk protein in the ovary of freshwater shrimp progene expression. Background technique [0002] Freshwater shrimp, scientific name Macrobrachium nipponense, commonly known as river shrimp, belongs to the order Decapoda, the family Palaemonidae, and the genus Macrobrachium. It is widely distributed in Asia in freshwater and low-salinity estuary areas. The country is one of the important ways to increase agricultural efficiency and farmers' income in our country. According to the 2016 "China Fishery Yearbook", the annual output value of farmed freshwater shrimp in the country is nearly 20 billion yuan, which is one of the most promising species in the freshwater aqua...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/435C12N15/89A01K67/033
CPCA01K67/0333A01K2227/70A01K2267/02C07K14/43509C12N15/89
Inventor 傅洪拓李菲乔慧张文宜蒋速飞熊贻伟龚永生金舒博吴滟
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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