Cloning and Expression
of Annexin V cDNA in E. coli
SUN Jian-Xin, SHEN Xiao-Chuan and WU Xiang-Fu*
( Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai
200031, China )
Abstract The cDNA encoding the mature annexin
V was isolated by using RT-PCR method from total RNAs of fresh human placenta.
The result of sequencing indicated that the sequence of isolated annexin V cDNA
was the same as the reported nucleotide sequence of annexin V. The annexin V
cDNA was cloned into expression plasmid pET24a(+) under T7 promoter and then
transformed into E. coli BL21(DE3). SDS-PAGE analysis revealed that
the human annexin V was highly expressed and accumulated up to 38% of total
bacterial proteins in soluble form after the induction by 1 mmol/L IPTG. The
purified human annexin V can significantly prolong activated partial
thromboplastin time (APTT).
Key words Annexin V; RT-PCR; gene
cloning; gene expression; anticoagulant activity
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