ABSTRACT
The highly virulent PRRSV isolate strain HN-1/06 was cultivated on Marc-145. To study the viral entry mechanisms, the GP5 gene of PRRSV isolate was amplified by RT-PCR and cloned into pcDNA3.0 to generate the expressing plasmid pcDNA-GP5. pcDNA-GP5 was transfected into 293T by the calcium phosphate precipitation method. Analysis of flow cytometry confirmed that the GP5 proteins were expressed in surface of the 293T cells. Then 293T cells were transfected with pcDNA-GP5, pHIT60 and pHIT111 plasmids to generate pseudotyping virus. The pseudotyping virus supernatant was harvested 48 hours post-transfection and was detected by Western blotting and infection assay. Western blotting indicated that the GP5 glycoproteins were incorporated into the retroviral pseudotyped virus. Infection assay showed that the pseudotyped virus infected 293T and Mark-145 cell. The pseudotyped virus could be used to further study infectious mechanism of PRRSV.
Subject(s)
Animals , Mice , Cell Line , Cloning, Molecular , Endothelial Cells , Cell Biology , Metabolism , Virology , Leukemia Virus, Murine , Genetics , Metabolism , Porcine respiratory and reproductive syndrome virus , Chemistry , Genetics , Recombinant Proteins , Genetics , Swine , Transfection , Viral Envelope Proteins , Genetics , Virion , Genetics , MetabolismABSTRACT
The events of terror activities, natural disasters and infectious disease epidemics in the 21st century in the USA alert an urgent need to establish a novel hospital emergency management model. The federal government has proposed specific requirements for hospital emergency management in terms of organization, commanding and coordinating, emergency preparedness, training and rehearsal, resource management, communication and information transmission, Most of the American hospitals have adopted the model of hospital emergency management planning and preparedness, including mainly, assigning specialized personnel to hospital emergency management; establishment of an emergency management committee; formulation of an overall preparedness protocol for emergent events; performing fragility analysis; strengthening the coordination with extramural organizations and seeking expertise support; carrying out emergency preparedness training and rehearsal; and regular review and evaluation of the management. The commanding system set up especially to deal with various hospital emergent events is universally applied in American hospitals.