Cloning and Expression
of Targeting CuZn-SOD to Central
Nervous System
HE Hua-Jun1,2,
SHI Hui-Juan1, QIU Cun-Ping2, WU Xiang-Fu2*,
YUAN Qin-Sheng1*
( 1State Key Laboratory of Bioreactor Engineering and Institute
of Biochemistry, East China University of Science and Technology, Shanghai 200237,
China; 2Shanghai Institute of Biochemistry, the Chinese
Academy of Sciences, Shanghai 200031, China )
Abstract
Oxygen-derived free radicals are thought to be involved in the pathogenesis of
a wide range of neurological disorders. Targeted delivery of CuZn-SOD to
neurons in central nervous system may have therapeutic value in such diseases.
The gene encoding human CuZn-SOD was fused to tetanus toxin fragment C geneto
construct a fusion gene, then it was cloned into prokaryotic expression vector
pET-22b(＋)
and baculovirus vector pFastBacHTb, and was expressed in E.coli and
Tn-5B1-4 cells, respectively. The recombinant fusion protein has a subunit
molecular mass of 68 kD and is recognized by both anti-CuZn-SOD and
anti-tetanustoxin antibody. By pyrogallol autooxidation assay, it is shown that
the CuZn-SOD moiety retains substantial enzymatic activity, where the TTC
moiety might deliver the fusion protein to neurons in central nervous system.
So, CuZn-SOD/TTC may be a useful agent for the targeted delivery of CuZn-SOD to
neurons.
Key words CuZn-SOD； tetanus toxin fragment C； fusion protein； retrograde axonal transport； baculovirus expression system
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