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Adenovirus vector and its application in preparing HCV cell and mouse models

A technology of adenovirus and recombinant adenovirus, applied in the biological field, can solve the problems of HCV vaccine evaluation, pathogenic mechanism, and research that cannot be used

Active Publication Date: 2015-07-08
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing HCV infectious mouse models are mainly immunodeficient human liver chimeric mice, which cannot be used for HCV vaccine evaluation and pathogenic mechanism research

Method used

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  • Adenovirus vector and its application in preparing HCV cell and mouse models
  • Adenovirus vector and its application in preparing HCV cell and mouse models
  • Adenovirus vector and its application in preparing HCV cell and mouse models

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the acquisition of HDAd-HCV recombinant adenoviral vector

[0038] The third-generation adenovirus vector HDAd-HCV, which recombines the HCV type 2a JFH1 genome, is to insert sequence 1 in the sequence listing into the adenovirus vector HDAd (Prolonged Peritoneal Gene Expression Using AHelper-Dependent Adenovirus.Peritoneal Dialysis International, 2009; 29:508-516 , the public can obtain from the Institute of Microbial Epidemiology, Academy of Military Medical Sciences of the Chinese People's Liberation Army) The vector obtained between the I-Ceu I and I-Sce I sites.

[0039] The specific preparation is as follows:

[0040] 1. Construction of pMD18T-HCV500-Ribo:

[0041] The pJFH1 plasmid (Production of Infectious Hepatitis C Virus in Tissue Culture from ACloned Viral Genome. Nat. Med, 2005; 11(7): 791-796, available to the public from the Institute of Microbial Epidemiology, Academy of Military Medical Sciences of the Chinese People's Liberation Army) as...

Embodiment 2

[0053] Example 2, Application of HDAd-HCV Recombinant Adenoviral Vector in Preparation of Cell Model and Mouse Model

[0054] I, the acquisition of HDAd-HCV recombinant adenovirus

[0055] After the vector HDAd-HCV and HDAd empty vector obtained by Example 1 were all linearized with Pme I, they were mixed with the helper virus AdNG163 (Prolonged Peritoneal Gene Expression Using A Helper-Dependent Adenovirus.Peritoneal Dialysis International, 2009; 29:508-516, public (available from the Institute of Microbial Epidemiology, Academy of Military Medical Sciences of the Chinese People's Liberation Army) co-transfected 293 cells stably expressing Cre recombinase to package and activate recombinant HDAd-HCV, and then continuously added AdNG163 helper virus, respectively, through 60mm cell culture dishes, 150mm cell culture dishes and 3L cell culture flasks were expanded and cultured (37°C 60rpm shake flask culture for 3 days), the cells were collected and lysed and then subjected to ...

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Abstract

The invention discloses an adenovirus vector and its application in preparing HCV (hepatitis C virus) cells and mouse models. The invention provides a recombinant adenovirus vector which is obtained by inserting nucleotides shown by the sequence 1 in the sequence table in the adenovirus vector HDAd (helper-dependent adenovirus). The experiments prove that the invention provides a third generation adenovirus vector of a recombinant HCV 2a type full-length genome, the prepared adenovirus can be used for preparing HCV whole-genome cell models or mouse models, and the obtained models can be used for drug screening of anti-HCV drugs, so as to be used for HCV vaccine evaluation and researches of the pathogenic mechanism.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an adenovirus vector and its application in preparing HCV cells and mouse models. Background technique [0002] Hepatitis C virus (HCV) is one of the important factors causing chronic hepatitis, liver cirrhosis and liver cancer, which seriously threatens human health. At present, there is no effective preventive vaccine against HCV, and the lack of HCV models, especially infectious models, also hinders the research and development of HCV vaccines and pathogenic mechanisms, thus increasing the difficulty of its prevention and control. The existing HCV infectious mouse models are mainly immunodeficient human liver chimeric mice, which cannot be used for HCV vaccine evaluation and pathogenic mechanism research. (Mercer DF, Schiller DE, Elliott JF, Douglas DN., Hao C, Rinfret A, Addison WR, Fischer KP, Churchill TA, Lakey JR, Tyrrell DL, Kneteman NM. Hepatitis Cvirus replication in mice...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/861C12N5/10C12Q1/02A01K67/027A61K49/00
Inventor 周育森周小军孙世惠赵光宇郭彦
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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