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Shrimp molting inhibitory hormone gene and its application in accelerating molting and growth of green shrimp

A technology of molting inhibitory hormone and green shrimp, applied in the fields of biotechnology and developmental regulation

Inactive Publication Date: 2017-09-05
FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the application in the study of important genes in freshwater shrimp has not been reported

Method used

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  • Shrimp molting inhibitory hormone gene and its application in accelerating molting and growth of green shrimp
  • Shrimp molting inhibitory hormone gene and its application in accelerating molting and growth of green shrimp

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Example 1: Obtaining the full-length cDNA of the MIH gene of freshwater shrimp

[0021] Extraction of total RNA: Select two mature freshwater shrimps, remove the eye stalk tissue with sterile dissecting scissors, clip the eye stalk pigment part in RNA preservation solution (TaKaRaBio Inc. Japan), rinse and pre-treat the freshwater shrimp eye stalk For RNA extraction, refer to the Takara Trizol kit for specific steps.

[0022] First-strand cDNA synthesis: The first-strand cDNA was obtained after reverse transcription with reference to the Takara M-MLV reverse transcription kit with the above total RNA.

[0023] Obtaining the full-length cDNA of MIH gene in freshwater shrimp:

[0024] According to Macrobrachium rosenbergii ( Macrobrachium rosenbergii , KC990939.1) the conserved region of MIH cDNA, two pairs of intermediate primers were designed, see SEQ ID NO: 6~9,

[0025] (middle F1: 5′-CCGGCATTCCTTTGTTTACCTG-3, middle R1: 5′-ATATTCTGGCGTGTGGTCCTG-3′;

[0026] midd...

Embodiment 2

[0031] Embodiment 2: Obtaining of dsRNA of the freshwater shrimp MIH gene

[0032] According to the cloned freshwater shrimp MIH gene (Accession number: KF878973), the NCBI online sequence analysis software ( http: / / www.ncbi.nlm.nih.gov / gorf / gorf.html ) to determine the position of the open reading frame of the MIH gene as 436-795bp. According to the above results, the online dsRNA primer design software ( http: / / www.flyrnai.org / cgi-bin / RNAi_find_primers.pl ), design dsRNA primers in the MIH open reading frame of freshwater shrimp, and add the T7 promoter sequence 5'-TAATACGACTCACTATAGGG-3' in front of each primer to form dsRNA synthesis primers, and the primer sequences are the upstream primers SEQ ID NO:2 and downstream primer SEQ ID NO:3. All primers were synthesized at Bosun Biotechnology (Shanghai) Co., Ltd.

[0033] Using the above-mentioned green shrimp dsRNA primers to carry out PCR amplification on the total cDNA of green shrimp (the conditions are the same as...

Embodiment 3

[0034] Example 3: Injection of the dsRNA of the green shrimp MIH gene on the development of the green shrimp molting

[0035] 1. Selection of Shrimp for Experiment

[0036] In order to study the silencing effect of the dsMIH gene, this study first conducted a one-time RNAi experiment to select 50 adult female shrimp with strong vitality, uniform individuals, and a body weight of about 1.50±0.35g, and divided them into two groups on average. One group was the dsRNA injection group, The other group is the DEPC water injection group (control group). Before the experiment, the snails were inflated in a 500L polyethylene plastic white bucket for 72 hours to adapt to the laboratory breeding environment, and the snails were fed once a day in the morning and evening.

[0037] The continuous RNAi experiment was designed according to the results of the one-time RNAi experiment. The shrimps used in the continuous RNAi experiment were the same batch of hatched juvenile shrimp hatched in ...

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Abstract

The invention discloses a green shrimp molting inhibitory hormone gene and its application in accelerating the green shrimp molting and growth. The green shrimp molting inhibitory hormone gene has a nucleic acid sequence as shown in SEQ ID NO:1. Using RNA interference technology, RNA interference primers were designed according to the open reading frame of the green shrimp molting inhibitory hormone gene sequence, and then the total cDNA of green shrimp was used as a template for PCR amplification to obtain a sequence as shown in SEQ ID NO: 4, represented by SEQ ID NO : 4 Carry out in vitro transcription and synthesize dsRNA, after dsRNA is injected into the pericardial cavity of freshwater shrimp, accelerate freshwater shrimp molting and growth. The RNAi method has little damage to freshwater shrimp, does not affect its normal feeding, habitat and mating, is convenient for long-term research, and has a significant effect. The invention provides new ideas and effective means for regulating the growth and development of freshwater shrimp and cultivating fine varieties of freshwater shrimp.

Description

technical field [0001] The invention relates to the fields of biotechnology and development regulation, in particular to a green shrimp molting inhibitory hormone gene and its application in accelerating the molting and growth of green shrimp. Background technique [0002] 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 farmed freshwater shrimp in the country exceeds 237,000 tons, and the annual output value exceeds 15 billion yuan. Freshwater shrimp farming has played a pivotal role in my country's aquaculture. In recent years, with the continuous expansion of large-scale farming and the continuous improvement of consumer demand, the key to the current genetics and br...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/16C12N15/113A01K67/033
Inventor 江丰伟傅洪拓乔慧张文宜金舒博熊贻伟蒋速飞龚永生
Owner FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
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