Screening of Genes
Encoding Nuclear Localization Signal in a Mouse Embryonic cDNA Library
WANG Ji-Shu, SUN Qiang, LI Rong, ZHOU Peng, HAN Hua*
( Department of Biochemistry and Molecular Biology, The Fourth Military
Medical University, Xi′an 710032, China )
Abstract A genetic
screening system was established previously for the cloning of gene fragments
encoding nuclear localization signals (NLS). In the current study, the system
was improved and its ability to distinguish nuclear and cytoplasm-localized
proteins was verified by inserting a known NLS-encoding gene. A mouse 10.5-d pc
embryonic cDNA library was then inserted into the cloning site of the screening
vector and 22 out of 104 clones were shown to be positive after screening
using the system. The cDNA inserts of some of these clones were sequenced, and
typical and untypical NLS were identified. The nuclear localizing activity of
the cDNA inserts was further verified using the GFP fusion protein system. So,
the system is useful in the cDNA library screening of genes encoding
NLS-bearing proteins. �お�
Key words nuclear localization signal；mouse；embryo；gene cloning；yeast
*Corresponding author：Tel,86-29-3374517；e-mail,[email protected]