Ank/Aγ-globin retrovirus constructs. (A)
The integrated single-copy provirus. The LTR sequences are shown as
hatched boxes, the ankyrin promoter as a gray box, exon sequences in
the γ-globin gene as dark boxes, and intron sequences as open boxes.
The asterisk indicates the presence of a 717-bp deletion in IVS2 of the
γ-globin gene. (B) The double-copy retrovirus vector
in the producer cell genome (Upper) and the integrated
provirus (Lower). The elements are identified as in
A. Note that during the conversion of the RNA genome to
DNA, the 3′ LTR, which includes the Ank/Aγ-globin gene,
is duplicated into the 5′ LTR. The positions of the restriction enzyme
sites for HindIII and EcoRV are shown.
HindIII digestion detects intact 5′ LTR sequences and
generates a specific 3′ junction fragment for each insertion.
EcoRV digestion detects intact 3′ LTR sequences and a
specific 5′ junction fragment for each insertion.