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Fig. 1. Characterization of the oskA87 and
osk187 alleles. (A) Schematic representation of
the oskar locus and the insertion points of the transposable elements
in the new oskar alleles. E1 through E4 represent the oskar
exons. The first exon contains a 15 nucleotide untranslated region. In
oskA87, a ZAM retrotransposable element is inserted 51 bp
upstream of the first intron, in reverse orientation relative to
oskar. In osk187, an I element is inserted 534 bp
upstream of the oskar transcription initiation site, in the same
orientation as oskar. (B,C) Northern blot and RT-PCR
analysis of oskar mRNA in the new oskar mutants. (B)
Northern blot: WT1 sample is RNA from wild-type (Oregon R) egg chambers of
stages 1 to 14, and WT2 from wild-type egg chambers of stages 1 to 7-8. The
eglRC12 sample represents ovaries whose development
arrested during oogenesis at stages similar to the new oskar mutants.
osk54 and osk84 are nonsense alleles
of oskar (Kim-Ha et al.,
1991). Only trace amounts of oskar mRNA are detected in
homozygous osk187 egg chambers and no oskar mRNA
is detected in osk187/Df(3R)pXT103 egg
chambers. (C) RT-PCR: in contrast to bcd (upper panel), no
oskar mRNA is detected by RT-PCR in
oskA87/Df(3R)pXT103 egg chambers
(lower panel, lane 2) while a 126 bp band is detected in a control
amplification from nosA10/Df(3R)pXT103 (lower
panel, lane 1), an allele of nanos originating from the same genetic
background as oskA87. Lanes 3: negative controls without
addition of template. (D) oskar RNA in situ hybridization in
osk187 and oskA87.
Fluorescently-labeled antisense oskar RNA probe was used to visualize
the presence of endogenous oskar RNA in stage 3-4 wild-type,
osk187/Df(3R)pXT103 and
oskA87/Df(3R)pXT103 mutant egg chambers.
Compared with wild type, considerably less or no oskar RNA is
detected in osk187/Df(3R)pXT103 and in
oskA87/Df(3R)pXT103 egg chambers,
respectively.
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