Modified live-attenuated vaccines (MLV) created by DNA shuffling against porcine reproductive and respiratory syndrome virus (PRRSV)
AssigneeVirginia Tech Intellectual Properties, Inc.
InventorsYanyan Ni, Yaowei Huang, Xiang-Jin Meng
Life of this patent
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Abstract
The present invention provides a novel infectious cDNA clone of porcine reproductive and respiratory syndrome virus (PRRSV), particularly for PRRSV strain VR2385; an improved DNA-launched reverse genetics system for PRRSV; infectious chimeric PRRSV viruses generated through DNA shuffling; modified live-attenuated virus vaccines (MLV) using DNA shuffled chimeric viruses; chimeric viral proteins produced through shuffled chimeric viruses; PRRSV antigens and subunit vaccines based on shuffled chimeric viral proteins; and method of producing broadly protective PRRSV vaccines using DNA shuffling techniques.
Claims16
An infectious chimeric PRRSV comprising proteins that are chimeric of a plurality of genetically distinct strains, wherein the chimeric proteins are GP2, GP3, GP4, GP5, and M.
A method for producing an infectious chimeric PRRSV, wherein the infectious chimeric PRRSV comprises proteins that are chimeric of a plurality of genetically distinct strains, wherein the chimeric proteins are GP2, GP3, GP4, GP5 and M, comprising:
- mixing deoxyribonucleic acid molecules derived from a plurality of various genetically distinct strains of PRRSV encoding GP2, GP3, GP4, GP5 and M;
- digesting the deoxyribonucleic acid molecules by using a nonspecific deoxy nuclease;
- extending the digested deoxyribonucleic acid molecules via polymerase chain reaction without adding primer;
- amplifying chimeric deoxyribonucleic acid molecules encoding GP2, GP3, GP4, GP5 and M;
- inserting the amplified chimeric deoxyribonucleic acid molecules into an infectious deoxyribonucleic acid clone of PRRSV;
- and
- infecting a host cell with the infectious deoxyribonucleic acid clone of PRRSV.
Dependencies are inferred from the claim text.