建立誘導式開啟心臟專一性基因的轉殖魚(1/2)
Date Issued
2005
Date
2005
Author(s)
蔡懷楨
DOI
932313B002058
Abstract
The tetracycline–controlled transcription system is well developed in many transgenic
organisms. Yet, this system has not been reported in zebrafish. In order to develop tet-on
regulatory system to allow the transgenic GFP to be expressed specifically in the
zebrafish heart, we use the zebrafish cmlc2 promoter to drive the
reverse-transactivator(rtTA). We constructed vectors which presented GFP after Dox
induction in zebrafish fibroblast cell line. Then, we applied this system in vivo.
Although fluorescence appeared slightly leaky, transgene showed that rtTA in the
transient embryos and drive GFP specific in the heart. After we got the germ-line F1
embryos, we found that the Dox dosage depended induction efficiency (10 µg/ml、
1µg/ml、100 ng/ml、10 ng/ml、and 1ng/ml). After 6 hr induction under treating with 1
µg/ml Dox, the rtTA-derived heart-specific GFP expression appeared in the transgenic
lines embryos at 72-hpf with a relative low leakiness. The higher GFP levels, the longer
induction time: the highest level was reached within 36~48 hr after induction. The
germ-line transmitted F2 from F1 crossing wild-type has the same tendency of
induction, and the F2 from F1 crossing F1 shows stronger GFP intensity of highest level
within 48~60 hr after induction These results were consistent with cell line induction.
The leaky GFP is also stronger than those of F1 and F2 from F1 crossing wild-type.
This is the first tet-on system developed in transgenic fish, which should be very useful
to study gene function at any stage whenever gene is induced.
Publisher
臺北市：國立臺灣大學分子與細胞生物學研究所
Type
report
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