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  4. 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)
 
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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)

Date Issued
2002-07-31
Date
2002-07-31
Author(s)
蔡懷楨
DOI
902313B002260
URI
http://ntur.lib.ntu.edu.tw//handle/246246/20552
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.
Subjects
cis-element
EMSA
GFP
myf-5
somite-specific expression
transgenic
zebrafish
Publisher
臺北市:國立臺灣大學漁業科學研究所
Type
journal article
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