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Nucleic acid molecule used for interfering macrobrachium rosenbergii male reproduction, and preparation method thereof

A technology for male reproduction and Macrobrachium rosenbergii, which is applied in the direction of DNA preparation, the use of vectors to introduce foreign genetic material, DNA / RNA fragments, etc., can solve the problems of large amount of RNA, difficulty in maintaining interference efficiency, high synthesis cost, etc., and achieve good technical means and methods, low-cost, high-efficiency gene silencing effects

Active Publication Date: 2015-07-01
ZHEJIANG WANLI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used small interfering RNA is difficult to maintain long-term interference efficiency in animal experiments, and the amount of RNA required for animal experiments is large, and the synthesis cost is too high

Method used

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  • Nucleic acid molecule used for interfering macrobrachium rosenbergii male reproduction, and preparation method thereof
  • Nucleic acid molecule used for interfering macrobrachium rosenbergii male reproduction, and preparation method thereof
  • Nucleic acid molecule used for interfering macrobrachium rosenbergii male reproduction, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Preparation of double-stranded RNA that interferes with the male reproduction of Macrobrachium rosenbergii

[0041] 1. Molecular cloning

[0042] The applicant discovered a gene fragment related to the male development of Macrobrachium rosenbergii in previous research, and designed forward primers and reverse primers with restriction endonuclease sites for PCR amplification reaction. The forward primer (Forward Primer, FR) sequence is:

[0043] 5'-GC TCTAGA ATAATGGCTCCCTTCTTCAA-3' (underline marked as the restriction site of XbaI endonuclease); reverse primer (Reverse Primer, RP) sequence is

[0044] 5'-CG GGATCC GCTCAATCATAAAAACGTCC-3' (underlined is the restriction site of BamHI endonuclease).

[0045] Using the cDNA of the male reproductive system tissue of Macrobrachium rosenbergii as a template, PCR amplification was carried out.

[0046] The PCR reaction system is:

[0047]

[0048]

[0049] Add water to a final volume of 25 μl.

[0050] T...

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Abstract

The invention aims at providing a ribonucleic acid (RNA) molecule used for interfering macrobrachium rosenbergii male reproduction, and a preparation method thereof. The double-stranded ribonucleic acid molecule used for interfering macrobrachium rosenbergii male reproduction has a nucleotide sequence of SEQ ID NO:1 or 3. With the method provided by the invention, a large amount of double-stranded RNAs can be rapidly prepared with low cost and high efficiency. The preparation method is simple to operate. Also, with the nucleotide fragment screened by the invention, good interference efficiency can be achieved, and macrobrachium rosenbergii male reproductive system development can be effectively interfered. With the double strand provided by the invention, long-term and high-efficiency gene silencing can be realized in shrimp animals, such that good technical means and method are provided for the researches of shrimp functional genes.

Description

technical field [0001] The invention belongs to the technical field of gene deletion and inactivation, and specifically relates to a ribonucleic acid molecule for interfering with the male reproduction of Macrobrachium rosenbergii, which can be used for RNA interference research related to the male reproduction of Macrobrachium rosenbergii. Background technique [0002] RNA interference (RNA interference, RNAi) technology, as an important means of gene function research, has been widely used to explore the field of gene function in shrimp and crab animals. RNA interference refers to the phenomenon of efficient and specific degradation of homologous mRNA induced by double-stranded RNA, which is highly conserved during evolution. The use of RNAi technology can specifically knock out or turn off the expression of specific genes, so this technology has been widely used to explore the field of gene function. [0003] At present, RNA interference technology mainly uses small inte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/70C12N15/10
Inventor 马文明曹荣荣杨蓉钱国英李东瑞杨劲树杨帆杨卫军
Owner ZHEJIANG WANLI UNIV
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