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目的 :对酪氨酸蛋白激酶受体EphB2胞外的配体结合区基因进行表达研究。方法 :编码EphB2胞外的配体结合区基因片段克隆于pBluescriptSK(- )质粒中 ,经全自动序列分析仪测序正确后 ,再克隆重组入表达载体pGEX4T 1,构建高效表达克隆pHT ,转化大肠杆菌DH5α表达。结果 :经IPTG诱导 5h ,蛋白表达即达高峰 ,SDS PAGE及凝胶密度扫描分析 ,表达出约41kD大小的蛋白 ,占菌体总蛋白的 35 %。结论 :该区基因片段的高效表达为进一步制备其抗体 ,研究其配体 受体的相互作用等工作打下基础
OBJECTIVE: To study the expression of extracellular ligand binding domain of tyrosine protein kinase receptor EphB2. METHODS: The gene fragment coding for the extracellular ligand binding region of EphB2 was cloned into pBluescriptSK (-) plasmid. After sequencing by automatic sequence analyzer, it was recombined into expression vector pGEX4T 1 to construct highly expressed clone pHT and transformed into E. coli DH5α expression. Results: After induced by IPTG for 5h, the protein expression peaked. SDS-PAGE and gel density scanning analysis showed that the protein was about 41kD, accounting for 35% of the total bacterial protein. Conclusion: The high expression of gene fragments in this area lay the foundation for the further preparation of their antibodies and their interaction with ligand receptors