|Title:||魚類肌肉調控蛋白MYF-5及MYOGENIN的生物特性：在胚胎早期發育中的角色及互動(2/2)||Authors:||蔡懷楨||Keywords:||cis-element;EMSA;GFP;myf-5;somite-specific expression;transgenic zebrafish||Issue Date:||31-Jul-2002||Publisher:||臺北市：國立臺灣大學漁業科學研究所||Abstract:||
Myf-5, a transcription factor that controls muscle differentiation, is expressed in
somites during early embryogenesis. However, gene regulation of myf-5 is poorly
understood and detailed functional analysis of the regulatory cis-elements is needed.
In zebrafish, the myf-5 upstream sequence from -82 to -62 (-82/-62) was fused with a
basal promoter and transferred to fertilized zebrafish eggs. The -82/-62 cassette
drove GFP reporter gene expression specifically in the somites. Moreover, GFP
signals were detected exclusively in the somites of 28-hpf embryos derived from eggs
injected with 5×(-82/-62)/CMV/EGFP, which contained 5 copies of the -82/-62
cassette inserted within CMV promoter/enhancer. Thus, the -82/-62 cassette has
dual functions: to enhance somite-specific expression and to repress non-specific
expression during the early development of zebrafish embryos. This cassette
sequence was highly conserved in mouse myf-5 and had the same function. Mutated
sequence analysis and electrophoretic mobility shift assay of -82/-62 cassette showed
that the -70/-62 sequence is the major element for controlling myf-5 specificity and for
forming the DNA-protein complex. However, the putative CCAAT-like box, located
at -66/-62, could not direct somite-specific expression. Therefore, the -70/-67
sequence is essential for controlling somite-specific expression and the CCAAT-like
box is essential for activating gene transcription.
|Appears in Collections:||漁業科學研究所|
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