spindlin1基因的原核表达及多克隆抗体制备

Prokaryotic expression of spindlin1 and the polyclonal antibody production

目的:明确人新基因spindlin1的组织表达谱、相互作用蛋白。方法与结果:应用PCR技术获得spindlin1的开放读框,并将其插入原核表达载体pGEX-2T,转化大肠杆菌BL21(DE3),经IPTG诱导后,融合蛋白GST-Spindlin1获得了可溶性表达。用SDS-PAGE胶上的目的条带做抗原制备了多克隆抗体,进行了抗体纯化。结论:spindlin1在OVCar3细胞中可能以二聚体形式存在,这一现象还有待于进一步研究。

Object-five： Spindlin1 is a protein of 237 amino-acid residues which is a member of spin/ssty gene family. Its function remains largely unknown. Methods and Results： Spindlin1 ORF was inserted into the expressive vector pGEX-2T. Confirmed by restriction enzymes and sequencing analysis, the recombinant plasmid was introduced into the Escherichia coli BL21（DE3）. Induced by 0.1 mmol/L IPTG, the fusion protein GST-Spindlin1 was expressed and the GST- Spindlin1 was soluble. Explored the band on the polyacrylamide gel as antigen, the polyclonal antibody was raised. After purification of the antibody, western blot was performed to access the validity. Conclusion： In OVC ar3 cell, dimmer of Spindlin may be formed, and further study is needed about it.