人牙本质磷蛋白功能片段的原核表达

目的本研究旨在利用分子生物学技术原核表达包含功能区的人DPP片段.方法首先构建带有人DPP基因5'端序列的GST融合蛋白表达质粒.经IPTG诱导表达,SDS-PAGE电泳.结果人牙本质磷蛋白基因5'端序列可在原核表达系统中进行不溶性表达,表达的融合蛋白分子量约为43KDa,进行初步纯化后,经SDS-PAGE证实,在预期位置出现了纯化蛋白条带.结论利用GST融合蛋白表达系统表达了人DPP功能片段.为进一步通过对人DPP功能片段的研究揭示DPP在牙齿发育和牙髓修复中的作用及其机制奠定基础.

Prokaryotic Expression of human dentin phosphoprotein domain

Objective To clone and express the human DPP domain containing DSS was cloned and expressed using genetic engineering in prokaryotic expression system.Methods The 504-bp long fragment of human DPP gene was isolated from plasmid pT-DPP-N and subcloned into a GST (fusion protein) expression vector pGEX-5X-1. Results SDS-PAGE assays yielded a roughly 43000 Da expressed protein at the expected mobility position, which presented in inclusion body. After lysed with urea solution, the expressed proteins were separated from bacteria lysates with a commercially supplied chromatography sepharose 4B. Conclusion Human DPP domain is cloned and expressed using GST(fusion protein) expression system, which will lay a foundation for further study on the role of DPP in tooth development and pulp restoration.