Development Of A Vaccinia Virus Expression System For Structural Proteins Of Aleutian Mink Disease Parvovirus

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Authors
Berry, Bradley
Advisor
Rev. Joseph Harrington
Marshall Bloom
John Addis
Editor
Date of Issue
1992-04-01
Subject Keywords
Publisher
Citation
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Title
Development Of A Vaccinia Virus Expression System For Structural Proteins Of Aleutian Mink Disease Parvovirus
Other Titles
Type
thesis
Description
Abstract
Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.Aleutian Mink Disease Parvovirus (ADV) belongs to the parvovirus family. Although ADV can be grown in tissue culture and has been cloned in procaryotic systems, a eucaryotic expression system was desired. This would allow the opportunity to examine viral synthesis in absence of ADV replication. I attempted to develop a eucaryotic cloning system using vaccinia virus to express the ADV virion protein genes. My goal was to clone a segment of a cDNA clone (pIB21), that coded for the open reading frame containing both virion protein-1 (VP1) and virion protein-2 (VP2) of ADV, into pSCll, a modified vaccinia virus co-insertion vector. I attempted this cloning procedure with clones constructed from segments of two different strains of the ADV virus, ADV-G and ADV-Utah-1. The recombinant vaccinia virus coinsertion vectors generated from these clones were used by members of Dr. Marshall Bloom's research team at Rocky Mountain Laboratories. They homologously recombined them with live vaccinia virus by transfection into cells previously infected with vaccinia virus. The vaccinia system was successful in expressing the ADV genes and synthesizing the viral proteins. It was later demonstrated that these proteins self-assembled into viral particles.
Sponsors
Degree Awarded
Bachelor's
Semester
Spring
Department
Life & Environmental Sciences