Cloning, Expression and
Tumor Suppression of Human Endostatin
HE Zhi-Yong1,2, CHEN Zhe-Yu1,3, QIU
Cun-Ping1, LI Biao1, ZHANG Wei-Jie2, WU
Xiang-Fu1*
( 1Shanghai Institute of Biochemistry, the Chinese Academy of
Sciences, Shanghai 200031, China； 2College of Life Science and
Biotechnology, Shanghai Jiao Tong University, Shanghai 200030, China； 3Department of neurobiology, Second
Military Medical University, Shanghai 200433, China )
Abstract Human
endostatin cDNA was cloned from total RNA of normal Chinese liver cell line L02
by RT-PCR. Endostatin DNA sequence encoded 184 amino acid residues. Five base
pairs and 3 amino acid residues are different from that reported, it may be due
to interspecies difference. The endostatin cDNA was inserted into the
pET-28a(+) containing T7 promoter. The recombinant plasmid was transformed the E.coli
BL21(DE3). Recombinant human endostatin was highly expressed as inclusion body
when the expression strain BL-ENDO was induced with 1 mmol/L IPTG.
Result of SDS-PAGE analysis revealed that recombinant human endostatin was
accounted for up to 25% of soluble protein in E.coli. Purified and
refolded recombinant human endostatin was active in inhibiting tumor growth and
metastasis.
Key words endostatin； E.coli； cloning； expression； tumor repression
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