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Efficient Isolation of Regulatory Sequences from Human Genome and BAC DNA
- Source :
- Biochemical and Biophysical Research Communications. 290:1079-1083
- Publication Year :
- 2002
- Publisher :
- Elsevier BV, 2002.
-
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.
- Subjects :
- Transcriptional Activation
Chromosomes, Artificial, Bacterial
DNA, Complementary
Pseudogene
Biophysics
Electrophoretic Mobility Shift Assay
P1-derived artificial chromosome
Regulatory Sequences, Nucleic Acid
Biology
Transfection
Polymerase Chain Reaction
Biochemistry
Cell Line
Genes, Reporter
Humans
Genomic library
Molecular Biology
Gene
Genetics
Apolipoprotein A-I
Genome, Human
DNA
Cell Biology
Genome project
DNA-Binding Proteins
DNA binding site
Multigene Family
Human genome
K562 Cells
In vitro recombination
HeLa Cells
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 290
- Database :
- OpenAIRE
- Journal :
- Biochemical and Biophysical Research Communications
- Accession number :
- edsair.doi.dedup.....13b44941aea2ca7a6723842be84631a4