Codon-optimized reading frames facilitate high-level expression of the HIV-1 minor proteins.
We have constructed reading frames for the HIV-1 YU-2 minor proteins Vpr, Vpu, Vif and Nef that are codon-optimized for high-level expression in mammalian cells. We show that, in the absence of the Rev/Rev-response element system, these codon-optimized reading frames result in greatly increased levels of expression of the corresponding proteins in cell culture systems when compared with the native reading frame. Northern blot analysis shows that the increase in expression found with the codon-optimized reading frames is largely owing to increased steady-state mRNA levels.
Anson
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DS
Department of Genetic Medicine, Children, Youth, and Women's Health Service, 72 King William Road, North Adelaide, Australia. donald.anson@adelaide.edu.au
Dunning
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KR
D015152:Blotting, Northern; D015153:Blotting, Western; D003062:Codon; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D015693:Gene Products, nef / Q000096:biosynthesis / Q000235:genetics; D016346:Gene Products, vif / Q000096:biosynthesis / Q000235:genetics; D016336:Gene Products, vpr / Q000096:biosynthesis / Q000235:genetics; D015754:Genes, nef; D016341:Genes, vif; D016334:Genes, vpr; D016338:Genes, vpu; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D054298:Human Immunodeficiency Virus Proteins; D012333:RNA, Messenger / Q000096:biosynthesis; D016364:Reading Frames; D054334:Viral Regulatory and Accessory Proteins / Q000096:biosynthesis* / Q000235:genetics; D054311:nef Gene Products, Human Immunodeficiency Virus; D054320:vif Gene Products, Human Immunodeficiency Virus; D054325:vpr Gene Products, Human Immunodeficiency Virus; D000086082:Viroporin Proteins
D003062:Codon; D015693:Gene Products, nef; D016346:Gene Products, vif; D016336:Gene Products, vpr; D054298:Human Immunodeficiency Virus Proteins; D012333:RNA, Messenger; D054334:Viral Regulatory and Accessory Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D054320:vif Gene Products, Human Immunodeficiency Virus; D054325:vpr Gene Products, Human Immunodeficiency Virus; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins