Iodination of herpesvirus nucleic acids.
A simple method is described for the iodination of herpes simplex virus (HSV) DNA. The procedure involved synthesis of 125-I-labeled 5-iodo-dCTP which was subsequently used as a precursor for the in vitro repair synthesis of HSV DNA. Synthesis of 5-iodo-dCTP and purification from oxidation and reduction reagents, buffer salts, unreacted dCTP and Na125-I was accomplished in a single chromatographic step. It was possible to prepare 125-I-labeled HSV DNA in vitro with specific activities exceeding 10-8 counts/min/mu-g. The DNA prepared by this method reassociated with DNA extracted from HSV-infected HEp-2 cells but not with HEp-2 cell DNA. Iodinated HSV DNA was susceptible to S-1-endonuclease digestion once denatured but was resistant to digestion in the native form. This method was used to synthesize 125-I-labeled ribo-CTP (5-iodo-CTP) which was used to prepare cytomegalovirus-specific complementary RNA. The method should be of value in the preparation of viral probes and for use in autoradiography of viral nucleic acids.
D001345:Autoradiography; D002294:Carcinoma, Squamous Cell; D002460:Cell Line; D004279:DNA, Viral / Q000096:biosynthesis / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000378:metabolism; D006801:Humans; D007455:Iodine / Q000378:metabolism; D007457:Iodine Radioisotopes; D007553:Isotope Labeling / Q000379:methods*; D007822:Laryngeal Neoplasms; D008168:Lung / Q000196:embryology; D009693:Nucleic Acid Hybridization; D009694:Nucleic Acid Precursors / Q000378:metabolism; D018139:Simplexvirus / Q000378:metabolism
D004279:DNA, Viral; D003854:Deoxyribonucleotides; D007457:Iodine Radioisotopes; D009694:Nucleic Acid Precursors; D007455:Iodine