Construction of Biotinylated Annexin V
for Detection of Apoptosis
SUN Jian-Xin, and WU Xiang-Fu
( Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai
200031,China )
CHEN Zhe-Yu1, YOU Zhen-Dong
( Second Military Medical University, Shanghai 200433, China
)
Abstract The biotinylated annexin V was
generated by genetic engineering method. The encoding region of annexin V was
fused with a gene coding for the carboxyl terminal 87 residues of Escherichia
coli biotin carboxyl carrier protein. The fused gene was expressed in E.
coli, and the annexin V was biotinylated in vivo. The
biotinylation efficiency was about 60% as detected by HPLC. The biotinylated
annexin V was purified by avidin affinity chromatography and used to detect the
apoptosis of the neurons induced by morphine.
Key Words Annexin
V; biotinylation; E. coli; apoptosis；
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