Non-coding RNA expression profile analysis method for echinococcus granulosus protoscolex derived exosomes and cyst fluid derived exosomes and non-coding RNA sequences

A technology of Echinococcus granulosus, analytical methods, applied in the fields of biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc.

Pending Publication Date: 2021-12-24
中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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Problems solved by technology

However, whether E. granulosus-derived exosomes contain lncRNAs and whether their miRNAs can target host genes and regulate immune responses has not been reported.

Method used

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  • Non-coding RNA expression profile analysis method for echinococcus granulosus protoscolex derived exosomes and cyst fluid derived exosomes and non-coding RNA sequences
  • Non-coding RNA expression profile analysis method for echinococcus granulosus protoscolex derived exosomes and cyst fluid derived exosomes and non-coding RNA sequences
  • Non-coding RNA expression profile analysis method for echinococcus granulosus protoscolex derived exosomes and cyst fluid derived exosomes and non-coding RNA sequences

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Embodiment 1

[0151] 1. Materials and methods

[0152] 1.1 Sample source

[0153] The livers were collected from sheep naturally infected with Echinococcus granulosus in Xinjiang, and delivered to the laboratory in cold chain.

[0154] 1.2 Experimental method

[0155] 1.2.1 Separation of Echinococcus granulosus protoscole and cyst fluid

[0156] The liver of sheep naturally infected with Echinococcus granulosus was collected. After the surface was wiped and disinfected with 75% ethanol, the Echinococcus was punctured with a syringe in a biological safety cabinet to absorb the cyst fluid. After natural sedimentation in a 50ml centrifuge tube, the supernatant was transferred to A new centrifuge tube was used to collect the cyst fluid (300ml); the precipitate was washed 5 times with normal saline containing streptomycin (100U / ml) and penicillin (100μg / ml) to collect the original scoliosis. The obtained protoscole suspension was aspirated, stained with 1% trypan blue and counted under a micr...

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Abstract

The invention discloses a non-coding RNA (Ribonucleic Acid) expression profile analysis method for echinococcus granulosus protoscolex derived exosomes and cyst fluid derived exosomes. The method comprises the following steps: 1) separating echinococcus granulosus protoscolex from cyst fluid; (2) culturing the echinococcus granulosus protoscolex in vitro; (3) separating theechinococcus granulosus protoscolex derived exosomes and the cyst fluid derived exosomes , and obtaining the protoscopes derived exosomes PSC-ELVs and the cystic fluid derived exosomes HF-ELVs; 4) carrying out transmission electron microscope analysis; 5)carrying out nano particle size tracking analysis; (6)carrying out Western blotting; (7) constructing a miRNA library and sequencing; 8) constructing a whole transcriptome sequencing library and performing sequencing analysis; and (9) constructing a ceRNA regulation network. In addition, the invention also discloses non-coding RNA of the echinococcus granulosus protoscolex derived exosomes and cyst fluid derived exosomes obtained by screening with the method. According to the invention, the non-coding RNA expression profiles in the echinococcus granulosus protoscolex derived exosomes and the cyst fluid derived exosomes are obtained for the first time internationally.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a method for analyzing the non-coding RNA expression profile of Echinococcus granulosus protoscole and cystic fluid-derived exosomes. In addition, the present invention relates to the non-coding RNA sequences of Echinococcus granulosus protoscole and cystic fluid-derived exosomes. Background technique [0002] Exosomes are a type of vesicle-like body with a diameter of 30-150 nm, which can carry biologically active substances such as lipids, proteins, and nucleic acids, and play an important role in the exchange of substances and information transmission between cells. Exosomes have a lipid molecular layer, which keeps their contents stable during long-distance transport, and transfers functional components such as proteins, lipids, DNA, and RNA between cells through membrane fusion. Play an important immunomodulatory role in the development process. Exosomes can inhibit th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6851C12N15/113
CPCC12Q1/6888C12Q1/6851C12Q2600/158C12Q2600/178C12Q2531/113C12Q2525/207C12Q2563/107
Inventor 沈玉娟张小凡曹建平曹胜魁张璟
Owner 中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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