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Fig. 2
Artificial assays of ET activity of mMTI promoter.
(A) ET vector pTME and an artificial ET vector pTME-Z48 in miniTol2 transposons. ITR-R and ITR-L, right and left inverted terminal repeats of miniTol2 transposon; SA, splice acceptor from carp β-actin gene; Exon, exon2 from the carp β-actin gene; mMTI Pro., mMTI minimal promoter; BGH polyA, bovine growth hormone polyadenylation sequence. The Z48 enhancer was subcloned into the pTME vector upstream of ITR-R. (B and C) Zebrafish embryos were injected with pTME or pTME-Z48 at the one-cell stage and images were captured under a fluorescent microscope Zeiss M205 at 20 hpf. (D) The midbrain-specific expression of EGFP in one representative embryo from (C) injected with pTME-Z48 is shown in a merged fashion. (E) The average percentage of embryos with ubiquitous or midbrain specific GFP fluorescence in three independent experiments. Total numbers of embryos injected with pTME or pTME-Z48 were counted at 20 hpf and scored for ubiquitous EGFP expression and specific EGFP expression in the midbrain. (F) qRT-PCR analysis of relative EGFP mRNA levels in injected embryos. Data are given as means ± standard deviation (n = 3). ** indicates P < 0.01 versus the pTME-injected embryos.