Screening of Genes
Related to the Expression of Human ε-globin Gene by Using Yeast One-hybrid
System
HUANG Jian, HOU Chun-Hui, QIAN Ruo-Lan*
( Institute of Biochemistry and Cell Biology, Shanghai Institute for
Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031,
China )
Abstract The
developmental stage-specific silencing of the human ε-globin gene during
embryonic period is controlled, in part, by a silencer (from -392 to -177 bp)
upstream of this gene. In our previous work, by using DNase I footprinting
assay, a major protected region from -278 to -235 bp within this silencer DNA
was identified. In order to isolate genes encoding proteins that bind to this
target element, a human fetal liver cDNA library was screened by using yeast
one hybrid system. The candidate clones were sequenced, and the ribosomal
protein L3 (RPL3) was preliminarily isolated. Electrophoretic mobility shift
assay (EMSA) and competition tests also demonstrated its binding activity to
the target element. These imply that RPL3 might play an important role in
silencing the human embryonic ε-globin gene expression through the interaction
with this silencer.
Key words human ε-globin gene； yeast one-hybrid system； silencer； RPL3
*Corresponding author： Tel, 86-21-64315030-2092； Fax, 86-21-64331090； e-mail, [email protected]