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Application of seawater oryzias latipes HSP90ab1 gene and application of encoded protein of seawater oryzias latipes HSP90ab1 gene

A gene coding, seawater technology, applied in application, gene therapy, genetic engineering, etc., can solve the problem of unclear influence of HSP90ab1, and achieve the effect of improving survival rate

Active Publication Date: 2020-12-08
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006]Currently, the effect of HSP90ab1 on NNV infection is still unclear, and there is no report on the application of HSP90ab1 to resist neuronecrosis in fish

Method used

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  • Application of seawater oryzias latipes HSP90ab1 gene and application of encoded protein of seawater oryzias latipes HSP90ab1 gene
  • Application of seawater oryzias latipes HSP90ab1 gene and application of encoded protein of seawater oryzias latipes HSP90ab1 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Screening and identification of MmHSP90ab1 gene and expression of its encoded protein

[0025] (1) Test material

[0026] Seawater medaka haploid embryonic stem cells hMMES1 were cultured in ESM4 medium at 28°C. Digest and subculture with 0.25% trypsin every two days.

[0027] (2) Test method

[0028] 2.1 RNA extraction and reverse transcription

[0029] Collect approximately 1 x 10 in a 6-well plate by trypsinization 6 Density (unit / mL) hMMES1 cells were centrifuged at 1000rpm and then lysed with 1mL lysate (Trizol, Ivitrogen), mixed with chloroform and centrifuged, then extracted with isopropanol to obtain RNA. Then, use NanoDrop 2000 to detect the concentration and OD of RNA 260 / 280 , to ensure the concentration and purity of RNA, and use agarose gel electrophoresis to detect the integrity of RNA. Finally, using gDNA Eraser PrimeScript TM The RT (TaKaRa) kit reverse-transcribed the extracted RNA to synthesize cDNA.

[0030] 2.2 Amplification of the...

Embodiment 2

[0038] The living experiment of embodiment 2 seawater medaka fish

[0039] In order to study the effect of His-MmHSP90ab1 recombinant protein on the infectivity of RGNNV, healthy seawater medaka larvae were used for in vivo experiments. The specific experimental steps are as follows:

[0040] (1) Test material

[0041] Red-spotted grouper neuronecrosis virus (RGNNV) was isolated from larvae juveniles suffering from neuronecrosis in Guangdong Province, and was enriched in LJB cells, which were stored at -80°C.

[0042] (2) Test method

[0043] Seawater medaka larvae test was divided into three groups: MmHSP90ab1 experimental group (MmHSP90abl group), His control group (His group) and negative control group (Negative Control). During the test, RGNNV (100TCID 50 ) and the purified His-MmHSP90ab1 recombinant protein or His protein were incubated at 4° C. for 4 h. Then, the MmHSP90ab1 experimental group (n=10) was injected intraperitoneally with RGNNV incubated with MmHSP90ab1 ...

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Abstract

The invention belongs to the technical field of biology, particularly relates to an application of a seawater oryzias latipes HSP90ab1 (MmHSP90ab1) gene and an application of an encoded protein of theseawater oryzias latipes HSP90ab1 (MmHSP90ab1) gene, aims to research and develop a new method for resisting nervous necrosis of fishes, and provides an MmHSP90ab1 gene which is separated and screened from seawater oryzias latipes, and the gene and the encoded protein thereof can be applied to resisting nervous necrosis of fishes. The RGNNV incubated by the MmHSP90ab1 protein is injected into thesea oryzias latipes, and compared with a control group (taking His as incubation protein) injected with the RGNNV, the survival rate of the sea oryzias latipes can be remarkably increased, which indicates that the MmHSP90ab1 protein can be used for resisting nervous necrosis of fishes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of the medaka HSP90ab1 gene and its coded protein. Background technique [0002] my country is the world's largest aquaculture country, and aquaculture is an important part of my country's agriculture and an important growth point of the current agricultural economy. Fish is an important aquaculture species in our country. However, with the increase of aquaculture species and the expansion of aquaculture scale, various infectious diseases continue to break out and become popular, which has caused huge economic losses to my country's aquaculture industry. [0003] Fish nerve necrosis virus (Nervous Necrosis Virus, NNV) is one of the most harmful pathogens to farmed fish in my country. Fish neuronecrosis caused by this virus is a second-class animal disease in my country. The fatality rate can reach 100%. NNV can be transmitted horizontally through water bo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/17A61K39/00A61P31/14C07K14/46C12N15/12C12N15/70A23K50/80A23K20/147
CPCA61K48/005A61K48/0008A61K38/1706A61K39/0007A61P31/14C07K14/461C12N15/70A23K50/80A23K20/147
Inventor 易梅生贾坤同李烽全
Owner SUN YAT SEN UNIV
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