Genome-wide infectious cell model of human papillomavirus type 16

A human papillomavirus and whole-genome technology, which is applied in the field of establishing a human papillomavirus type 16 whole-genome infectious cell model, can solve the problems of long experimental period, hindering in-depth research on HPV, and complicated technology, and achieves a strong vitality. Effect

Active Publication Date: 2019-01-25
SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In conventional monolayer cell culture, the cells are in a poorly differentiated state, so the in vitro amplification of HPV cannot be achieved, which has hindered people's in-depth research on HPV for quite a period of time
The HPV nude mouse xenotransplantation model that appeared in the 1980s has promoted the research of HPV to a certain extent, but there are obvious defects such as long experimental period and complicated technology.
At present, there are no reports in this field in China

Method used

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  • Genome-wide infectious cell model of human papillomavirus type 16
  • Genome-wide infectious cell model of human papillomavirus type 16
  • Genome-wide infectious cell model of human papillomavirus type 16

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0033] Example Construction of pcDNA3.3-HPV16 infectious cloning plasmid.

[0034] According to the whole genome sequence of HPV16 strain with accession number U89348.1 in Genbank, 4 pairs of PCR amplification primers were designed and synthesized. Extract viral genomic DNA from HPV16-infected cervical squamous epithelial cells, use it as a PCR template, and use 4 pairs of primers for PCR amplification to obtain 4 DNA fragments, connect the 4 DNA fragments to T vectors, and select positive Cloned and sequenced. After confirming that the clone was correct, the four fragments were connected to the pcDNA3.3 vector in the order of BamHI-HPV16-BamHI using LIC cloning technology to obtain the recombinant pcDNA3.3-HPV16 cloning plasmid. After resistance screening, positive clones were selected and sequenced.

[0035] Using the SeqMan software in DNAStar, the measured sequences were spliced ​​to obtain the full-length viral genome sequence. And use the MegAlign software in DNAStar to ...

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Abstract

The invention relates to the technical field of bioengineering, in particular to the establishment of a genome-wide infectious cell model of human papillomavirus type 16. The whole genome of human papillomavirus type 16 (HPV16) in Northeast China was cloned and sequenced. The cyclic HPV16 genome was transfected into cells to establish a cell model containing the whole genome of HPV16 in NortheastChina. The invention establishes an in vitro simulation system capable of fully representing HPV-infected human host cells, and converts the system into an in vitro pharmacodynamic model of an anti-HPV16 drug.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the establishment of human papillomavirus type 16 whole genome infectious cell model. Background technique [0002] Human papillomavirus (HPV) is a small non-enveloped, double-stranded circular DNA virus. Its typing is based on the sequence similarity of the L1 gene. The sequence similarity of the L1 gene is <90% as a new type. , the similarity between 90% and 98% is different subtypes of the same type, and the similarity >98% is different variant strains of the same type. [0003] Human papillomavirus (HPV) is an epitheliophilic small DNA virus, which is the pathogenic microorganism of various wart diseases in humans. In recent years, it has been found to be related to various malignant tumors. The HPV virus is composed of a core closed-loop double-stranded DNA and a peripheral protein capsid. The genome is about 7.9kb in length and consists of 8 main open reading ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/37C12N15/85
CPCC12N15/85C07K14/005C12N2710/20022C12N2510/00
Inventor 吴盈盈路一平吴思王爽于淼刘淼姚雪春卢星忠
Owner SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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