Expression of additional genes in a vector derived from a minimal RNA virus.
Previous studies have shown that expression of the vesicular stomatitis virus (VSV) glycoprotein (G) from a Semliki Forest virus (SFV) RNA replicon results in the production of propagating infectious particles that we call minimal viruses. These minimal viruses consist of vesicles containing VSV G protein that bud from the plasma membrane and trap the infectious SFV G RNA, but they do not contain other viral structural proteins. The cell binding and membrane fusion activity of the VSV G protein allow minimal viruses to propagate in tissue culture cells. To determine if these minimal viruses could be used to express foreign genes, we added a second SFV promoter and a multiple cloning site downstream of the VSV G gene. We report here expression of three different proteins from this modified, minimal virus vector. Although expression of each foreign, unselected gene was lost rapidly from the vector upon passaging, it was possible after the initial transfection to derive stocks of infectious particles that could be used to infect multiple additional cultures and transfer protein expression efficiently. When cells were infected with these minimal viruses, host protein synthesis was shut off and the foreign protein and VSV G proteins were the major proteins expressed in the infected cells. Both were expressed at similar levels and accumulated to about 1-2% of total cell protein.
Rolls
M M
MM
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Haglund
K
K
Rose
J K
JK
D000818:Animals; D001483:Base Sequence; D015704:CD4 Antigens / Q000096:biosynthesis / Q000235:genetics; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D006224:Cricetinae; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012672:Semliki forest virus / Q000235:genetics*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics
D015704:CD4 Antigens; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; D015500:Chloramphenicol O-Acetyltransferase