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Recombinant adenovirus expressing human rotavirus VP7 and its preparing process and usage

A technology of recombinant adenovirus and rotavirus, which is applied in the field of medical genetic engineering, can solve the problems of immune difficult infection protection, achieve good immune effect, prevent or reduce invasion, and ensure the effect of safety and reliability

Inactive Publication Date: 2002-11-27
王健伟 +3
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

However, the genes of animal viruses are quite different from those of humans, and the immunity induced by them is difficult to use for human infection protection

Method used

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  • Recombinant adenovirus expressing human rotavirus VP7 and its preparing process and usage
  • Recombinant adenovirus expressing human rotavirus VP7 and its preparing process and usage
  • Recombinant adenovirus expressing human rotavirus VP7 and its preparing process and usage

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Embodiment Construction

[0030] 1. Cloning of human rotavirus VP7 gene

[0031] (1) Extraction of rotavirus genomic RNA (dsRNA)

[0032] Diarrhea stool samples were collected from the pediatric outpatient clinic of a domestic hospital, and confirmed to be rotavirus infection by electron microscopy. Take the supernatant of 10% stool sample diluted in PBS, after SDS treatment, phenol, chloroform extraction, ethanol precipitation, centrifugation, obtain rotavirus genomic RNA, and dissolve it in deionized water;

[0033] (2) Purification of rotavirus RNA

[0034] Viral nucleic acid was purified with the Nucleo Trap Gel Extraction Kit produced by CLONTECH Company according to the method provided by the product. The purified nucleic acid was dissolved in deionized water, placed at room temperature for 30 minutes, centrifuged, and the supernatant was frozen at -20°C;

[0035] (3) Amplification of VP7 cDNA

[0036]Using purified rotavirus RNA as a template, the entire VP7 gene was amplified by reverse tran...

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Abstract

A recombinant adenovirus expressing human rotavirus VP7 and its preparing process and application are disclosed. It is a non-replicative recombinant one with deficient regions E1 and E3 in its genom.The human rotavirus VP7 gene expression box is inserted into the position where there is deficient region E1. It can infect the intestine tract and the respiratory tract, induce the generation of mucosa immune and neutralize the antibody. It can be used to preven the acute infantal diarrhea caused by rotavirus infection.

Description

(1) Technical field [0001] The invention relates to a recombinant adenovirus expressing human rotavirus VP7, especially a recombinant adenovirus expressing infant rotavirus VP7, which belongs to the field of medical genetic engineering. [0002] The invention also relates to the preparation method and application of the recombinant adenovirus. (2) Background technology [0003] Rotavirus is the most important cause of diarrhea in infants and young children. It is widely prevalent in the world and causes huge losses every year. According to the report of the World Health Organization, in Asia, Africa, and Latin America, there were 30 cases of diarrhea in 1977-1978. ~5 billion cases of diarrhea occurred, and 5 to 10 million children died of diarrhea. Diarrhea ranked first in terms of morbidity and mortality. Another report estimated that in Asian, African, and Latin American countries (excluding China) , there are 740 million to 1 billion cases of diarrhea every year, and 4.6...

Claims

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Application Information

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IPC IPC(8): A61K39/235C12N7/01C12N15/33C12N15/63C12N15/861
Inventor 王健伟何金生洪涛徐倏燊
Owner 王健伟
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