Expression of r-PA and Kringle2 in E. coli and The Interactions with β2GPI
Fragments of r-PA and Kringle2 were obtained by PCR and then recombinant plasmids, pQE30-rPA and pRSETa-K2, were constructed and transformed into E. coli strain BL21(DE3), respectively. The expression of the target proteins was induced by 1 mmol/L IPTG and was identified by SDS-PAGE. The result showed that vast majority of the proteins was expressed in the format of inclusion body. Kringle2 and r-PA proteins were purified by immobilized metal ion-affinity chromatography (IMAC) from the dissolved inclusion bodies. The fibrinolytic activities of r-PA and Kringle2 were evaluated in vitro by fibrin agarose plate assay. We found that the catalytic activity of r-PA was enhanced when the concentration of β2GPI increased, whereas Kringle2 not. β2GPI was found in enzyme-linked immunosorbent assay (ELISA) to bind specially with r-PA and Kringle2, respectively.
t-PA β2GPI expression and purification ELISA
Shi-Jing Sun Chun-E Zhang Guo-Ping Cai
Life Science and Ocean Biology Laboratory of Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China
国际会议
The 5th International Forum on Post-genome Technologies(5IFPT)(第五届国际后基因组生命科学技术学术论坛)
苏州
英文
2007-09-10（万方平台首次上网日期，不代表论文的发表时间）