Cosolvents facilitate DNA synthesis in the herpes simplex virus 1 unique short (Us) inverted repeat.
DNA synthesis under standard conditions is not successful within a portion of the Us1 gene of HSV-1 which is juxtaposed to an 86% G + C-containing tract in the Us inverted repeat sequence. We report that the independent addition of specific amounts of at least three different types of cosolvents is capable of facilitating DNA synthesis within this G + C-rich region. In addition, this strategy was used to successfully place a specific site-directed mutation in the Us1 gene. Consideration of these observations should enable future site-specific mutational analyses of portions of the HSV-1 genome which have traditionally been unamenable to genetic manipulations.
Paragas
J
J
Department of Microbiology, Mount Sinai School of Medicine, New York, NY 10029-6574, USA.
Blaho
J A
JA
D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000096:biosynthesis*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004996:Ethidium; D005559:Formamides / Q000494:pharmacology; D005990:Glycerol / Q000494:pharmacology; D018259:Herpesvirus 1, Human / Q000235:genetics*; D017874:Immediate-Early Proteins / Q000235:genetics*; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D017830:Octoxynol; D017354:Point Mutation; D011092:Polyethylene Glycols / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012997:Solvents / Q000494:pharmacology; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
D004279:DNA, Viral; D005559:Formamides; C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D012997:Solvents; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C078424:EUS1 protein, Equine herpesvirus 1; D011092:Polyethylene Glycols; C031066:formamide; D017830:Octoxynol; C010615:Nonidet P-40; D004262:DNA Restriction Enzymes; D004996:Ethidium; D005990:Glycerol; D004121:Dimethyl Sulfoxide