Rhinovirus Vector for ER Stress Modeling
Endoplasmic reticulum (ER), as a new place to induce apoptosis in cells, can mediate ER stress (ERS) reactive apoptosis pathway.
Rhinovirus (RhV) is the most important pathogen causing the common cold. In recent years, a large number of studies have shown that RhV can not only cause upper respiratory tract infection, but also closely related to lower respiratory tract diseases such as bronchiolitis, pneumonia and asthma. RhV can also co infect with other viruses and bacteria to aggravate asthma and chronic obstructive pulmonary disease.
The nonstructural protein 2B of rhinovirus has the characteristics of low relative molecular weight and high degree of hydrophobic region. It can integrate into cellular ER and cause cellular pathophysiological changes, which may play an important role in the pathogenesis of RhV.
Using the above characteristics, Creative Bioarray constructed the eukaryotic expression vector of rhinovirus nonstructural protein 2B. By transfecting the expression vector, the endoplasmic reticulum stress model can be established in host cells.
Technology Principle
The nonstructural protein 2B of rhinovirus has the characteristics of low relative molecular weight and high degree of hydrophobic region, and can be integrated into ER cells. In order to more directly the effect of reaction modeling, we added green protein label on the carrier.
We transfected BHK-21 cell line with recombinant plasmid and identified it by Western blot.
Service Content
- Construction, amplification and marketing of plasmids
- Host cell culture and plasmid transfection services
- Construction of ERS model
In order to identify whether the constructed plasmid can normally express rhv16 nonstructural protein 2B in cells, we will transfect the plasmid into host cells after double enzyme digestion and sequencing, collect cells at a certain time after transfection, and check the expression of 2B fusion protein by Western blot.
In addition, we will locate the target protein to ensure that nonstructural protein 2B is expressed on ER.
In order to ensure the successful induction of ERS, we will detect the marker proteins GRP78 and chop of ERS after transfection.
- Endoplasmic reticulum stress model modeling effect and subsequent index detection service
Model Application
The transfected host cells can be used as a model of endoplasmic reticulum stress injury.
The construction of the plasmid not only lays a foundation for clarifying the cell damage mechanism of nonstructural protein 2B, but also provides a new potential target for the design of antiviral drugs.
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