TY - JOUR
T1 - Identification of 5′-upstream region of pufferfish ribosomal protein L29 gene as a strong constitutive promoter to drive GFP expression in zebrafish
AU - Chang, Ming Huang
AU - Chou, Chih Ming
AU - Hsieh, Yueh Chun
AU - Lu, I. Ching
AU - Devi, M. Kothai Nachiar
AU - Chang, Juei Pei
AU - Kuo, Tzong Fu
AU - Huang, Chang Jen
N1 - Funding Information:
We thank Dr. P. W. Hsiao for critically reading the manuscript and helpful discussion. This research was supported by grants from the National Science Council (NSC-91-3112-P-001-018-Y-3), Taiwan, Republic of China.
PY - 2004/1/30
Y1 - 2004/1/30
N2 - The genomic structure of Tetraodon fluviatilis L29 gene was determined and its promoter activity was analyzed in COS-1 cells and zebrafish embryos. The TfL29 gene comprises four exons and three introns, spanning approximately 1.7kb. The 5′-upstream 2.2-kb of the first exon contains 10 E-boxes and many putative binding motifs for transcription factors GATA-1, AML-1a, c-Myb, Oct-1, CdxA, and NRF-2. Promoter activity assay showed that the distal 2.2-kb fragment not only had high luciferase activity in COS-1 cells, but also strong and ubiquitous GFP expression in a variety of tissues in zebrafish embryos. On the other hand, there are no TATA or CAAT boxes within a 300-bp region upstream from the transcription initiation site. Although this region has high luciferase activity in COS-1 cells, it is not sufficient to drive GFP expression in zebrafish embryos. In this proximal 300-bp region, there are two E-boxes, two CdxA sites, and one NRF-2 site that is immediately downstream of the transcription start site.
AB - The genomic structure of Tetraodon fluviatilis L29 gene was determined and its promoter activity was analyzed in COS-1 cells and zebrafish embryos. The TfL29 gene comprises four exons and three introns, spanning approximately 1.7kb. The 5′-upstream 2.2-kb of the first exon contains 10 E-boxes and many putative binding motifs for transcription factors GATA-1, AML-1a, c-Myb, Oct-1, CdxA, and NRF-2. Promoter activity assay showed that the distal 2.2-kb fragment not only had high luciferase activity in COS-1 cells, but also strong and ubiquitous GFP expression in a variety of tissues in zebrafish embryos. On the other hand, there are no TATA or CAAT boxes within a 300-bp region upstream from the transcription initiation site. Although this region has high luciferase activity in COS-1 cells, it is not sufficient to drive GFP expression in zebrafish embryos. In this proximal 300-bp region, there are two E-boxes, two CdxA sites, and one NRF-2 site that is immediately downstream of the transcription start site.
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U2 - 10.1016/j.bbrc.2003.12.080
DO - 10.1016/j.bbrc.2003.12.080
M3 - Article
C2 - 14715273
AN - SCOPUS:0347356239
SN - 0006-291X
VL - 314
SP - 249
EP - 258
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -