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Transfection system of taking eimeria tenella virus ribonucleic acid (RNA) as carrier

A technology of Eimeria, transfection system, applied in the field of transfection system, nucleic acid sequence of Eimeria tenella virus

Inactive Publication Date: 2014-05-07
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It has been reported in Eimeria tenella, Eimeria stezeri, Eimeria nisslovakia, Eimeria brucei, Eimeria maxima, Eimeria venomousa, and heap-type Eimeria and other in vivo electron microscopes have observed virus-like particles, but the nucleic acid sequence of Eimeria tenella virus and the virus-based transfection system have not been reported yet

Method used

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  • Transfection system of taking eimeria tenella virus ribonucleic acid (RNA) as carrier
  • Transfection system of taking eimeria tenella virus ribonucleic acid (RNA) as carrier

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Embodiment 1

[0016] Transfection system using Eimeria tenella virus RNA as carrier

[0017] as attached figure 1 As shown, according to the sequence characteristics of Eimeria tenella virus (EtRV1) genome dsRNA, the green fluorescent protein (enhanced green fluorescent protein, EGFP) gene was replaced with part of the dsRNA coding region to construct the chimera of EtRV1 and GFP. The nucleic acid sequence of Eimeria tenella virus is shown in SEQ ID NO:1. Using PCR reaction on EtRV1 dsRNA to introduce the T7 promoter into the 5′ end of dsRNA cDNA, using PCR method to amplify EtRV1’s 5′ end 377bp (including 5′ end UTRs), 3′ end 141bp, EGFP coding region 717bp three A fragment was constructed to construct a chimera of EtRV1 and EGFP, and cloned into the vector pMD-18T respectively to obtain a recombinant plasmid pMD-18T-5'UTR-EGFP-3'UTR, which was linearized by EcoRI and HindII? Gel recovery kit for recovery and quantification. About 2μg of the linearized plasmid was used as a template, a...

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Abstract

The invention provides a transfection system of taking an eimeria tenella virus ribonucleic acid (RNA) as a carrier. The transfection system contains polyclonal restriction sites, and is convenient to use, and a plurality of exogenous genes can be switched into the eimeria tenella body, so as to carry out research of gene expression regulation, biochemical metabolism, worm development and the like of the eimeria tenella.

Description

[0001] Technical field: [0002] The invention provides a transfection system using Eimeria tenella virus RNA as a carrier, and discloses the nucleic acid sequence of Eimeria tenella virus at the same time, belonging to the field of genetic engineering. [0003] Background technique: [0004] Coccidiosis (Coccidiosis) is caused by Eimeria coccidiosis, and the damage is very serious parasitic disease. As an intracellular parasitic eukaryote, Eimeria has caused huge losses in the global poultry industry. Although it has been studied for many years, there is no ideal control method so far. The reason is that the coccidian cells and molecular organisms lack of detailed understanding. Double-stranded RNA (dsRNA) viruses were first discovered in fungi in 1948, and dsRNA viruses have since been found in many protozoa. It has been reported in Eimeria tenella, Eimeria stezeri, Eimeria nisslovakia, Eimeria brucei, Eimeria maxima, Eimeria venomousa, and heap-type Virus-like particle...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/40C12N15/86C12R1/93
Inventor 李建华张西臣武斌宫鹏涛信彩岩张国才张楠杨举李赫杨正涛
Owner JILIN UNIV