Restriction digests of the clone give the following sizes (kb): EcoRI/BamHI--2.7, 2.1, 1.65 (triplet); EcoRI--2.7, 2.1, 1.65 (triplet); PstI--4.0, 2.4, 1.3 (doublet), 0.5; BglI--2.5, 1.65 (doublet), 1.5, 1.4, 1.1; BglII--6.5, 1.65 (doublet). Restriction digests of the clone give the following sizes (kb): BamHI/EcoRI--2.7, 2.1, 1.7 (triplet); PstI--4.0, 2.5, 1.3 (doublet), 0.50. The unit genome contains the following restriction sites (bp from the 5' end): BglI--716; BglII--224; EcoRI--1427; HincII--962, 1200; NcoI--576; PstI--651, 1083; SalI--962; SmaI--479, 1107; XbaI--781; XhoI--1272, 1506, 1545. Excising with BamHI=EcoRI yields 3 insert fragments of 1.7 kb and vector fragments of 1.8 kb and 3.0 kb--the construct has a unique BamHI site. Virus DNA cloned after passage in liver of woodchuck The insert consists of a head to tail trimer of the full length genome (1.679 kb) beginning at the unique EcoRI site and oriented so that transcription from the SV40 late promoter gives genomic RNA. Excising with BamHI+EcoRI yields 3 insert fragments of 1.7 kb and vector fragments of 1.8 kb and 3.0 kb. The construct has a unique BamHI site. Transfected COS-7 cells replicate the HDV RNA genome, and produce the 195 aa small form of the delta antigen, encoded from nt 1598 to 1014. |
