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RNAi reagent for interfering portunus trituberculatus sex determining gene expression as well as RNAi method and application thereof

A technology of portunus trituberculatus and gene expression, which is applied in the direction of DNA/RNA fragments, other methods of inserting foreign genetic materials, genetic engineering, etc., can solve the problems of unsuitable aquatic larvae, individual damage, individual size, etc., to achieve small damage and improve The effect of success rate and broad application prospect

Pending Publication Date: 2022-02-08
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

However, because RNAi mostly adopts methods such as injection, which has a certain degree of damage to the individual and has certain requirements for the individual size, it is not suitable for aquatic larvae

Method used

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  • RNAi reagent for interfering portunus trituberculatus sex determining gene expression as well as RNAi method and application thereof

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Experimental program
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Embodiment 1

[0030] 1. Preparation of RNAi reagents

[0031] The sequence is shown in SEQ ID No.1 Dmrt The gene sequence is sent to Sangon Bioengineering (Shanghai) Co., Ltd. for primer design and preparation into RNAi reagents, which contain a sense strand (Sense) and an antisense strand (Antisense) forming a double-stranded region , the sequence is as follows:

[0032] Sense: GCGCAUCCAUACUAACAAATT (SEQ ID No. 2);

[0033] Antisense: UUUGUUAGUAUGGAUGCGCTT (SEQ ID No. 3).

[0034] 2. Soak rotifers with RNAi reagent

[0035] Dilute the prepared RNAi reagent with DEPC water to a concentration of 3.3 μg / ml, soak the rotifers for 2-3 hours, and prepare for feeding.

[0036] 3. Feeding of Portunus trituberculatus

[0037] The Portunus trituberculatus used in the present invention all come from the experimental base of Changyi Haifeng Aquatic Products Co., Ltd., Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences. The female crabs were placed in a culture pond (50...

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Abstract

The invention provides an RNAi reagent for interfering portunus trituberculatus sex determining gene expression as well as an RNAi method and application thereof. The RNAi reagent comprises a sense strand and an antisense strand which form a double-strand region; and the sequence of the sense strand is as shown in SEQ ID No.2, and the sequence of the antisense strand is as shown in SEQ ID No.3. The RNAi reagent can be used for efficiently knocking down the expression of a Dmrt gene in the early development period of the portunus trituberculatus, damage to the portunus trituberculatus is small, then the RNAi reagent can be applied to sex determination mechanism analysis and sex control technology research of the portunus trituberculatus, the method is easy to operate, breakthrough of the unisexual breeding technology of the portunus trituberculatus is facilitated, and thus wide application prospects are realized.

Description

technical field [0001] The invention belongs to the technical field of gene interference, and in particular relates to an RNAi reagent for interfering with the expression of a sex-determining gene of Portunus trituberculatus and its RNAi method and application. Background technique [0002] Most crustaceans are sexually dimorphic and have differences between males and females. The realization of parthenogenesis can significantly improve economic benefits. At present, decapod economical crustaceans can only be knocked down in the larval stage by knocking down the expression of key sex-determining genes in Macrobrachium rosenbergii. And realize. [0003] Portunus trituberculatus ( Portunus trituberculatus ) belongs to Crustacea, Decapod order, Portunidae, commonly known as Portunus, is an important large-scale marine economic crab in my country. Studies have shown that the Dmrt gene is an important sex-determining gene in Portunus trituberculatus, but there is no knockdown m...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/87A01K67/033C12Q1/6851
CPCC12N15/113C12N15/87A01K67/0338C12Q1/6851C12N2310/14A01K2207/05A01K2227/70A01K2267/02C12Q2531/113C12Q2545/101C12Q2563/107Y02A40/81
Inventor 吕建建张文高保全刘萍李健
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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