The insert contains internal EcoRI, AvaI, XbaI, SalI, AccI, HincII and PstI sites. The internal XbaI site is methylated in HB101 and will not cut if host is dam+. Restriction digests of the clone give the following sizes (kb): BamHI--2.9, 2.7; EcoRI--5.4, 0.11 (doublet); HindIII--5.6; PstI--3.5, 2.2; XbaI--5.6. Linearize the plasmid with HindIII for in vitro transcription. This is a full-length clone of virus segment S4. The cDNA ends were tailored to terminal BamHI sites before cloning into the BamHI site of pGEM-1. |
