Journal: Journal of Virology
Article Title: Mutational Analysis, Using a Full-Length Rubella Virus cDNA Clone, of Rubella Virus E1 Transmembrane and Cytoplasmic Domains Required for Virus Release
Figure Lengend Snippet: Construction of RV M33 virus full-length cDNA clone. The numbers on the viral genome scale refer to the distance from the 5′ end in kilobases. Six DNA fragments were amplified by proofreading TaKaRa Taq DNA polymerase with the requisite primers as described in Materials and Methods. The amplified DNA fragments were ligated into a full-length cDNA representing the whole viral genome by using the restriction sites indicated above the genome. The full-length cDNA was cut with Eco RI and Hin dIII and inserted into a modified pBR322 plasmid that had been cut with the same enzymes to obtain the full-length cDNA clone pBRM33.
Article Snippet: PCR amplifications were performed in separate reactions containing cDNA, 1 pmol of primers, 0.4 mM deoxynucleoside triphosphates, 10% dimethyl sulfoxide, and proofreading TaKaRa Taq polymerase (Takara Shuzo Co., Ltd.) in a buffer provided by the manufacturer.
Techniques: Amplification, Modification, Plasmid Preparation