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Biotechniques 1989 Oct;7(9):980-982

Improved retroviral vectors for gene transfer and expression.

Miller AD, Rosman GJ

Fred Hutchinson Cancer Research Center, Seattle, WA 98104.

We describe a set of murine retrovirus-based vectors that include unique cloning sites for insertion of cDNAs such that the cDNA can be driven by either the retroviral long terminal repeat, the immediate early promoter of human cytomegalovirus, or the simian virus 40 early promoter. The vectors carry the neomycin phosphotransferase gene expressed from an alternate promoter as a selectable marker. These vectors have been constructed to prevent viral protein synthesis from the remaining viral sequences, to yield high-titer virus stocks after introduction into retrovirus packaging cells, and to eliminate homologous overlap with viral DNAs present in retrovirus packaging cells in order to prevent helper virus production. Methods for generating high-titer virus are described.

MeSH Terms:

• Animal
• Biotechnology
• Cell Line
• Gene Expression Regulation, Viral
• Genetic Markers
• Genetic Vectors*
• Helper Viruses/physiology
• Helper Viruses/genetics
• Retroviridae/physiology
• Retroviridae/genetics*
• Support, U.S. Gov't, P.H.S.
• Transfection
• Viral Proteins/genetics
• Viral Proteins/biosynthesis
• Virus Replication

Substances:

• Viral Proteins
• Genetic Markers

Secondary source id:

• GENBANK/M28248
• GENBANK/M28247
• GENBANK/M28246
• GENBANK/M28245

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