人骨涎蛋白在大肠杆菌中的表达

Expression of human bone sialoprotein in E.Coli

目的 :在大肠杆菌中表达骨涎蛋白 (BSP)。方法 :构建BSP -PBV2 2 0表达载体 ,重组子以大肠杆菌DH5α为宿主菌进行诱导表达。结果 :工程菌经 4h 42℃热诱导后 ,在SDS -PAGE电泳上出现了一条新的蛋白带 ,Mr为 33× 10 3 ～ 35× 10 3 。结论 :BSP在大肠杆菌中得到了表达 ,其Mr为 33× 10 3 ～ 35× 10 3 ,为进一步蛋白纯化及抗体制备奠定了基础。

AIM: To express the human bone sialoprotein(BSP) in E.Coli. METHODS: 900bp cDNA fragment of BSP was obtained with EcoRⅠand PstⅠfrom the plasmid pBS-BSP. The fragment was inserted into expression vector PBV220. The recombinant plasmid PBV-BSP was transformed into E.coli DH5α and expressed by means of heat-induction at 42℃ for 4h. RESULTS: SDS-PAGE revealed a new foreign protein band near 33×10 3～35×10 3(Mr). CONCLUSION: BSP protein was obtained. The expression of BSP will make it possible for further study.