[Construction and functional identification of red fluorescent protein reporter plasmid for human catalase gene].
OBJECTIVE
To construct a red fluorescent protein reporter gene driven by human catalase gene promoter.


METHODS
The red fluorescent protein reporter gene plasmid pDsRed-CATp containing human catalase gene promoter was constructed by gene recombination technique. The plasmid was transiently transfected into NIH/3T3 cells to observe their response to H(2)O(2) stimulation.


RESULTS
The plasmid was constructed correctly as verified by double enzyme digestion and sequence analysis. The plasmid was lowly expressed in resting NIH/3T3 cells, but the expression level increased obviously after stimulation by H(2)O(2). CONCLSIONS: A red fluorescent protein reporter gene plasmid driven by human catalase gene promoter has been constructed successfully with a sensitive response to H(2)O(2) stimulation. This system provides a convenient tool for the study of the regulatory mechanism of catalase gene expression.
Yao
Qi
Q
Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou 510515, China.
Huang
Shao
S
Liu
Ya-Wei
YW
Liu
Jing-Hua
JH
Jiang
Yong
Y
D016475:3T3 Cells; D000818:Animals; D002374:Catalase / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics*; D005822:Genetic Vectors / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014162:Transfection; D000097573:Red Fluorescent Protein
D002374:Catalase; D006861:Hydrogen Peroxide; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein