Skip to main content
. 2001 Aug;75(15):7030–7041. doi: 10.1128/JVI.75.15.7030-7041.2001

FIG. 2.

FIG. 2

Regulation of expression of the enzymatically active antiviral Gag-SN fusion protein in transgenic lines (A) Time course of expression of the antiviral transgene in testis. Testis tissues from 34- to 343-day-old line E and F mice were characterized by immunoblot analysis with anti-SN antibody. Results were reproduced with two or three mice for each age and line. Virus particles containing Gag-SN fusion proteins (virions) (36) served as positive controls. Testis tissue from nontransgenic littermates of line E and F mice, respectively, served as a negative control (−C). The other bands migrating faster than the 85-kDa Gag-SN fusion protein are caused by nonspecific cross-reaction. (B) Antiviral gene products are detectable in lymphatic tissue of line E mice. LN tissue from line F (age 145 days) and line E (age 75 days) mice as well as C57BL/6 (negative control [−C]) mice was analyzed. One additional band in line E tissue in the 40-kDa range (arrow) may result from proteolytic processing of the 85-kDa Gag-SN fusion protein. (C) RT-PCR analysis of poly(A) RNA from B and CD90+ T cells isolated from LN confirms the presence of gag-SN transcripts (546-bp RT-PCR product) in lymphatic cells of line E and F mice. The animals were 412 (E1), 371 (E2), and 460 (F [−C]) days old. PCR without cDNA synthesis (−RT) served as a control for contamination of the poly(A) RNA preparation with genomic DNA. Poly(A) RNA from a nontransgenic littermate (−C) was used as negative control. (D) Zymogram analysis of testis tissue expressing Gag-SN fusion proteins demonstrates nucleolytic activity of the fusion protein. The gel was loaded with 2 mg of testis tissue extract from transgenic lines E and F, a nontransgenic littermate (−C), and C57BL/6. Gag-SN incorporating MuLV particles that were released from tissue culture cells (virions) served as a positive control for nucleolytic activity. Gag-SN precursor protein and products of proteolytic processing incorporated into virus particles are depicted as boxes. (E) Immunoblot analysis of viral pellet fractions from sera of line E and F mice with anti-SN antibody. VLP preparations from 23 homozygous mice each of lines E and F are shown. VLP preparations from 23 nontransgenic C57BL/6 mice served as negative control (−C). Gag-SN incorporating MuLV particles (virions) released from tissue culture cells (36) and 5 ng of purified SN were loaded as positive controls.