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Isolated fish antiviral protein gene CMPK2 and antiviral activity thereof

An anti-virus and protein technology, applied in anti-viral agents, gene therapy, genetic engineering, etc., can solve the problems that the leucine zipper domain has not been found and needs further research

Active Publication Date: 2019-03-08
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More than 60% of leucine, alanine, proline and glycine in CMPK2 are located in the N-terminal domain, but no leucine zipper domain was found, and contrary to the leucine-rich nature, in this There is no isoleucine in the domain, so the leucine-rich N-terminal domain may have special properties, such as protein-protein interactions, which will be further studied

Method used

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  • Isolated fish antiviral protein gene CMPK2 and antiviral activity thereof
  • Isolated fish antiviral protein gene CMPK2 and antiviral activity thereof
  • Isolated fish antiviral protein gene CMPK2 and antiviral activity thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0019] (1) RNA extraction from FHM cells

[0020] Plate the FHM in good condition on a 6-well plate, discard the supernatant after 24 hours, add 1ml TRIzol reagent, and place it in a 1.5ml RNase-free EP tube.

[0021] Add 200ul of chloroform solution, vortex to mix, and let stand on ice for 5 minutes.

[0022] Centrifuge at 10,000 x g for 15 minutes at 4°C. The sample is divided into three layers: the bottom layer is an organic phase, the upper layer is a colorless aqueous phase and an intermediate layer.

[0023] Transfer the aqueous phase to a new tube, add an equal volume of isopropanol, point to mix, let stand on ice for 10 minutes, and centrifuge at 10,000×g for 10 minutes at 4°C. Remove the supernatant and add 75% ethanol to wash the precipitate.

[0024] Centrifuge at 10,000×g for 5 minutes at 4°C, add an appropriate amount of RNase-free water to the centrifuge tube to dissolve the RNA, and set aside at -80°C.

[0025] (2) Reverse transcription to obtain total cDNA ...

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Abstract

The invention belongs to the technical field of genetic engineering of aquatics and particularly relates to isolated fish antiviral protein gene CMPK2 and antiviral activity thereof. An antiviral natural immunoprotein gene CMPK2 is isolated from Pimephales promelas; a nucleotide sequence of this gene is shown as the sequence table SEQ ID NO: 1; an encoded protein sequence of this gene is shown asSEQ ID NO: 2. The expression of the protein gene CMPK2 cloned herein may affect proliferation of SVCV (spring viremia of carp virus); CMPK2 protein plays an important role in the life cycle of SVCV. The gene or protein herein provides a new target for the research on the preparation of anti-SVCV drugs.

Description

technical field [0001] The invention belongs to the technical field of freshwater fish genetic engineering, and in particular relates to an isolated fish antiviral protein gene CMPK2 and identification of antiviral activity thereof. Background technique [0002] Carp spring viremia is an important aquatic animal disease caused by carp spring viremia virus (SVCV). In my country, the Ministry of Agriculture also lists it as a second-class animal disease. SVCV is a rhabdovirus. Now listed as a provisional member of the Rhabdoviridae, genus Vesicular Stomatosis. The virus infection has a high mortality rate and causes huge economic losses. Once the fry are infected with the disease, they show as slow swimming, low excitability, staying at the bottom of the pond or gathering in the water, unresponsive and until death. Diseased fish mainly manifested as internal hemorrhage, peritonitis, and visceral hemorrhage spots in appearance. Meanwhile, SVCV can infect herring, grass car...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12C12N15/54A61K48/00A61K38/45A61P31/14
CPCA61K38/45A61K48/005A61P31/14C12N9/1205C12Y207/01074
Inventor 刘学芹刘皖蒙刘焦云姚健王业大王方陈博
Owner HUAZHONG AGRI UNIV
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