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Application of STCH to preparation of functional product for resisting flavivirus infection

A technology of virus infection and flavivirus, applied in the field of biomedicine, can solve problems such as infection and achieve good application prospects

Active Publication Date: 2021-08-06
大汉生物科技(广东)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no effective drug for preventing and treating flavivirus infection. In order to solve the harm caused by flavivirus infection and transmission, it is urgent to develop a Novel effective agent or drug against flavivirus infection

Method used

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  • Application of STCH to preparation of functional product for resisting flavivirus infection
  • Application of STCH to preparation of functional product for resisting flavivirus infection
  • Application of STCH to preparation of functional product for resisting flavivirus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Determination of STCH gene transcription level after flavivirus infection of A549 cells

[0030] 1. Collection of test samples:

[0031] Total RNA samples of A549 cells infected with ZIKV, DENV, JEV and YFV were collected.

[0032] 2. The detection method specifically includes the following steps:

[0033] (1) A549 cells according to 2×10 5 cells / ml in 12-well plates and cultured overnight.

[0034] (2) Discard the culture medium the next day and wash twice with 1×PBS. Take DENV infection as an example: Infect cells at MOI=1, absorb for 1 hour, and shake well every 15 minutes. After 1h, discard the adsorption solution, wash twice with FBS-free DMEM, add 1ml maintenance solution, and incubate for 48h.

[0035] (3) Total cellular RNA was collected at 0h, 24h and 48h, and Quantitative Real-time PCR (Q-RT-PCR) was used to detect the transcription level of STCH gene.

[0036] 3. Experimental results:

[0037] The result is as figure 1 The results showed tha...

Embodiment 2

[0038] Example 2 Determination experiment of flavivirus gene transcription level after overexpressing STCH in A549 cells

[0039] 1. Collection of test samples:

[0040] Total RNA samples of A549 cells infected with DENV, JEV, YFV and ZIKV after overexpression of STCH were collected.

[0041] 2. The detection method specifically includes the following steps:

[0042] (1) A549 cells according to 2×10 5 cells / ml in 12-well plates and cultured overnight.

[0043] (2) Discard the culture medium the next day and wash twice with 1×PBS. Taking DENV infection as an example: an HA-tagged STCH overexpression plasmid (HA-STCH, lug / ul) was constructed, transfected into A549 cells, and then the cells were infected with the virus at MOI=1, and cultured for 24 hours.

[0044] (3) Total cellular RNA was collected, and the transcription level of the virus was detected by Q-RT-PCR.

[0045] 3. Experimental results:

[0046] The result is as figure 2 It was shown that the gene transcriptio...

Embodiment 3

[0047] Example 3 Determination of Flavivirus Protein Transcription and Gene Translation Levels After Overexpression of STCH in A549 Cells

[0048] 1. Collection of test samples:

[0049] The total RNA samples and total protein samples of A549 cells infected with ZIKV and DENV after overexpression of STCH were collected.

[0050] 2. The detection method specifically includes the following steps:

[0051] (1) A549 cells according to 2×10 5 cells / ml in 12-well plates and cultured overnight.

[0052] (2) Discard the culture medium the next day and wash twice with 1×PBS. Taking DENV infection as an example: an HA-tagged STCH overexpression plasmid (HA-STCH, lug / ul) was constructed, transfected into A549 cells, and then the cells were infected with the virus at MOI=1, and cultured for 24 hours.

[0053] (3) Total cellular RNA and total protein were collected, and the transcription and translation levels of viral proteins were detected by Q-RT-PCR and Western Blot.

[0054] 3. E...

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Abstract

The invention belongs to the field of biological medicine, and discloses an application of a stress chaperone molecule (STCH) to preparation of a functional product for resisting flaviviruses. By flavivirus inhibition experiments, the invention proves that an STCH overexpression vector has a remarkable inhibition effect on the flaviviruses, which shows that the STCH overexpression vector is expected to prevent and treat diseases caused by a flavivirus infection when being applied to preparation of a drug for resisting the flavivirus infection.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and more specifically relates to the application of STCH in the preparation of functional products resistant to flavivirus infection. Background technique [0002] Flavivirus (Flavivirus) belongs to the Flavivirus genus of the family Flaviviridae. It is a large group of enveloped single positive-strand RNA viruses with a diameter of 40-70 nm and a icosahedral stereosymmetry. This type of virus causes infection through the transmission of blood-sucking arthropods (mosquitoes, ticks, sandflies, etc.), and was classified as arboviruses in the past. At present, those that have a greater impact on humans include Dengue Virus (DENV), Zika Virus (ZIKV), Japanese Encephalitis Virus (JEV), West Nile Virus (WNV) ) and yellow fever virus (Yellow Fever Virus, YFV) and so on. [0003] Diseases caused by dengue virus (DENV) infection include: general fever, mild dengue fever (dengueFever, DF), severe den...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K48/00A61P31/14
CPCA61K38/1709A61K48/005A61P31/14Y02A50/30
Inventor 黄曦王巧花吴永坚
Owner 大汉生物科技(广东)有限公司
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