Efficient isolation of regulatory sequences from human genome and BAC DNA.
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.
Xu
Hai-Ming
HM
National Laboratory of Medical Molecular Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100005, People's Republic of China.
Zhang
Shen
S
Liu
De-Pei
DP
Li
Xing-Guo
XG
Hao
De-Long
DL
Liang
Chih-Chuan
CC
D016632:Apolipoprotein A-I / Q000235:genetics; D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial; D004247:DNA / Q000302:isolation & purification*; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D017930:Genes, Reporter; D015894:Genome, Human; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D012045:Regulatory Sequences, Nucleic Acid; D015533:Transcriptional Activation; D014162:Transfection
D016632:Apolipoprotein A-I; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D004247:DNA