[Application of EGFP as reporter gene in study of WT1 regulation elements].
In order to investigate the feasibility of using EGFP as a reporter gene in WT1 transcriptional regulation study. WT1 promoter and enhancer were ligated into the vector pEGFP-1 by recombinant DNA technique and confirmed by restriction enzymes digestion. The resultant constructs were transfected into K562 cell line by DMRIE-C reagent and the function of these WT1 gene elements was detected by using a fluorescent microscope after transfection for 48 hours. The results indicated that the recombinant vectors, pEWP containing WT1 promoter, and pEWPE, pEWPA and pEWPD harboring both WT1 enhancer and promoter, had been successfully constructed. Fluorescence was observed in K562 cells transfected by pEWP, pEWPE, pEWPA and pEWPD, while no fluorescence could be detected in cells transfected by pEGFP-1. It is concluded that EGFP gene as a reporter gene can be applied to the WT1 transcriptional regulation study, which provides the basis for gene therapy.
Hu
Shao-Yan
SY
Jiangsu Institute of Hematology, The First Affiliated Hospital of Suzhou University, Suzhou 215006, China.
Chen
Zi-Xing
ZX
Zhao
Ye
Y
Fu
Zheng-Zheng
ZZ
He
Jun
J
Cen
Jian-Nnong
JN
Gu
Min
M
D004742:Enhancer Elements, Genetic / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D014162:Transfection; D025721:WT1 Proteins / Q000235:genetics*
D025721:WT1 Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins