Fig. 1.
Fig. 1. pinvVIIILA transgene construct. / From 5′ to 3′: human involucrin promoter region (including 1.2-kilobase intron), SV40 intron sequences, human factor VIIILA (B domain–deleted) cDNA, and SV40 polyadenylation signal. The domain composition of factor VIIILA, including the retained remnant of the central B domain (ΔB), is depicted. The transgene is liberated from the vector backbone by SalI digestion prior to zygote injections. The 5′ and 3′ primers used for RT-PCR analysis flank the intron sequences as shown. The upstream primer is based in 5′ untranslated sequences derived from the involucrin promoter region, while the downstream primer is derived from factor VIII coding region. Therefore, the 210–base pair RT-PCR amplification product from this primer pair is specific for fully spliced, transgene-derived message. The transcription initiation site is designated by the arrow upstream of the first intron.

pinvVIIILA transgene construct.

From 5′ to 3′: human involucrin promoter region (including 1.2-kilobase intron), SV40 intron sequences, human factor VIIILA (B domain–deleted) cDNA, and SV40 polyadenylation signal. The domain composition of factor VIIILA, including the retained remnant of the central B domain (ΔB), is depicted. The transgene is liberated from the vector backbone by SalI digestion prior to zygote injections. The 5′ and 3′ primers used for RT-PCR analysis flank the intron sequences as shown. The upstream primer is based in 5′ untranslated sequences derived from the involucrin promoter region, while the downstream primer is derived from factor VIII coding region. Therefore, the 210–base pair RT-PCR amplification product from this primer pair is specific for fully spliced, transgene-derived message. The transcription initiation site is designated by the arrow upstream of the first intron.

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