Construction and functional identification of red fluorescent protein reporter plasmid for human catalase gene / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To construct a red fluorescent protein reporter gene driven by human catalase gene promoter.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p>