Efficient isolation of regulatory sequences from human genome and BAC DNA.
Biochem Biophys Res Commun, 2002/1/25;290(3):1079-83.
Xu HM[1], Zhang S, Liu DP, Li XG, Hao DL, Liang CC
Affiliations
PMID: 11798185
Impact factor: 3.322
Abstract
Isolation of regulatory DNA fragments is the basis of the identification of DNA binding proteins and the study of the regulation of gene expression. Presently there is a lack of efficient methods to broadly isolate and identify DNA regulatory fragments. We developed an efficient method to isolate regulatory DNA sequences from both genome and bacterial artificial chromosome (BAC) based on electrophoretic mobility shift assay and PCR techniques without purified transcription factors. Twenty-nine DNA fragments were isolated from human genome and 24 from BAC DNA containing human apolipoprotein AI gene cluster. Transient transfection assay showed that some fragments could enhance the transcription of reporter gene.
MeSH terms
Apolipoprotein A-I; Cell Line; Chromosomes, Artificial, Bacterial; DNA; DNA, Complementary; DNA-Binding Proteins; Electrophoretic Mobility Shift Assay; Genes, Reporter; Genome, Human; HeLa Cells; Humans; K562 Cells; Multigene Family; Polymerase Chain Reaction; Regulatory Sequences, Nucleic Acid; Transcriptional Activation; Transfection
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