P-选择素凝集样区和绿色荧光蛋白融合基因的构建

Construction of P-selectin lectin-like domain fused with green fluorescent protein gene

目的 构建P 选择素凝集样区与绿色荧光蛋白融合基因 (pEGFP N1/L) ,为进一步明确P 选择素不同表位与功能间的关系 ,为阐明P 选择素生理病理意义奠定基础。方法 应用PCR技术扩增质粒pCDM 1/cDNA凝集素样区 ,用T4连接酶将其插入载体 pEGFP N1;经酶切和测序对插入片段进行分析、鉴定。 结果 通过酶切和序列分析证实插入片段序列正确。结论 应用基因工程技术构建 pEGFP N1/L融合基因成功 ,为绿色荧光蛋白作为生物标记分子来研究P

Objective To construct a fused gene of P selectin lectin like domain and green fluorescent protein(pEGFP N1/L) to provide the scientific basis to clarify the pathological physiology sense of P selectin. It also has laid a foundation to analyze the relationship between molecular structure and function. Methods The lectin like region fragment of P selectin obtained by PCR was insert into pEGFP N1. The fusion gene was verified by enzyme digestive analysis and sequencing. Results A positive band was formed when product of PCR electrophoresis on agarose gel and insert fragment was confirmed correctly by enzyme digestion analysis and sequencing. Conclusion The fusion gene pEGFP N1/L was constructed successfully by genetic engineer technique. It has laid a foundation to study P selectin's structure function relationship by GFP used as biology marker molecular.