Expression, Purification
and in vitro Activity of Apoptin
SUN Guo-Jing, TONG Xin, DONG Yan, SUN Zhi-Xian*
( Institute of Radiation Medicine, Academy of Military Medical Sciences,
Beijing 100850, China )
Abstract A novel
protein named apoptin induces a p53-independent, Bcl-2 insensitive type
apoptosis in various human tumor cells but not in normal cells. Apoptin is
considered to be a promising candidate for safe and effective
anti-tumortherapy. Moreover, apoptin may be important for fundamental studies
on molecularbasis of apoptosis and cancer. Apoptin gene was subcloned into
prokaryotic expression vector pPROEXHT and pBV220, respectively. The apoptin
protein was obtained from pPROEXHT-apoptin expression system and not from
pBV220-apoptin； the former contains a 6×histidine
affinity tag for ease of purification. Expressed protein was purified by
chromatography on a co-chelated affinity column and was renatured by dialysis.
The renatured apoptin was able to induce purified Hela nuclei apoptosis in
vitro even without the participation of the cytoplasm, showing that
apoptin expressed in E.coli was active to induce apoptosis.
Key words apoptin； prokaryotic expression； purification； induce apoptosis
*Corresponding author： Tel, 86-10-66931218； Fax, 86-10-68214653； e-mail, [email protected]