Restriction digests of the clone give the following sizes (kb): SacI--4.0; HindIII--4.0; SalI--4.0; PstI--3.3, 0.7. The insert contains the following restriction sites (nt position according the published sequence): KpnI--249, 674, 1008; PstI--454; SacI--506; NruI--883. Linearizing with SphI or HindIII and transcription with SP6 RNA polymerase will give the sense strand. The SP6-derived transcript would direct the translation of the V2 (coat protein) ORF in an in vitro translation system, such as wheat germ or rabbit reticulocyte lysate. Constructed from the MluI/SpeI fragment extending from nt 183 to 1071 of the published sequence. The EcoRI/MluI and SpeI/XbaI sites were destroyed. |
